15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022].

See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
THE ANATOMICAL RECORD PART A 276A:75–102 (2004)

Adult Stem Cells

HENRY E. YOUNG1,2* AND ASA C. BLACK, JR.1,3
1
Division of Basic Medical Sciences, Mercer University School of Medicine,
Macon, Georgia
2
Department of Pediatrics, Mercer University School of Medicine, Macon, Georgia
3
Department of Obstetrics and Gynecology, Mercer University School of Medicine,
Macon, Georgia

ABSTRACT
Development of a multicellular organism is accomplished through a series of events that
are preprogrammed in the genome. These events encompass cellular proliferation, lineage
commitment, lineage progression, lineage expression, cellular inhibition, and regulated ap-
optosis. The sequential progression of cells through these events results in the formation of
the differentiated cells, tissues, and organs that constitute an individual. Although most cells
progress through this sequence during development, a few cells leave the developmental
continuum to become reserve precursor cells. The reserve precursor cells are involved in the
continual maintenance and repair of the tissues and organs throughout the life span of the
individual. Until recently it was generally assumed that the precursor cells in postnatal
individuals were limited to lineage-committed progenitor cells specific for various tissues.
However, studies by Young, his colleagues, and others have demonstrated the presence of two
categories of precursor cells that reside within the organs and tissues of postnatal animals.
These two categories of precursor cells are lineage-committed (multipotent, tripotent, bipo-
tent, and unipotent) progenitor cells and lineage-uncommitted pluripotent (epiblastic-like,
ectodermal, mesodermal, and endodermal) stem cells. These reserve precursor cells provide
for the continual maintenance and repair of the organism after birth. Anat Rec Part A 276A:
75–102, 2004. © 2004 Wiley-Liss, Inc.

Key words: adult; pluripotent; stem cells; mammals; humans; embryonic; mes-
enchymal; epiblastic

The formation of tissues and organs occurs naturally dur-
ing embryogenesis (review, Moore and Persaud, 1998; Carl-
son, 1999; Kacsoh, 2000). Development of a multicellular
organism follows multiple predetermined molecular and cel-
lular pathways that culminate in the formation of an indi-
vidual composed of billions of cells with well-defined respon-
sibilities. These responsibilities include provision for
structural support and mobility, electrical and chemical con-
trol of body functions, protection through controlled interac-
tions with the surrounding environment, nutrition, removal
of waste products, and procreation. Moreover, the cells, tis-
sues, and organs must be maintained throughout the life
span of the individual as well as repaired following trauma
or disease. Cellular development is accomplished through
the genome as a series of preprogrammed events. These
events include cellular proliferation, lineage commitment,
lineage progression, lineage expression, cellular inhibition,
and regulated apoptosis. Sequential progression through
these programmed events results in the formation of the
differentiated cells, tissues, and organs that fashion an indi-
vidual. These preprogrammed processes are initiated follow-
ing fusion of the male and female pronuclei to form a totipo-
tent zygote (Fig. 1).
As development proceeds from the single-celled zygote,
proliferation of the blastomeres forms a solid sphere,
termed the morula. Further proliferation and lineage com-
mitment results in formation of a blastocyst, a hollow
sphere of cells composed of the trophoblast, germ cells,
and the inner cell mass. The trophoblast continues to
develop by proliferation and segregation into the cytotro-
phoblast and syncytiotrophoblast. These two layers even-
tually form the embryonic portion of the placenta (Fig. 1).
The germ cells, precursors to oogonia (female) or sper-
Grant sponsor: Rubye Ryle Smith Charitable Trust; Grant
sponsor: MEDCEN Community Health Foundation; Grant spon-
sor: MorphoGen Pharmaceuticals, Inc.
*Correspondence to: Dr. Henry E. Young, Division of Basic
Medical Science, Mercer University School of Medicine, 1550
College St., Macon, GA 31207. Fax: 478-301-5489.
E-mail: young_he@mercer.edu
Received 27 January 2003; Accepted 21 February 2003
DOI 10.1002/ar.a.10134
Published online 00 Month 2004 in Wiley InterScience
(www.interscience.wiley.com).

© 2004 WILEY-LISS, INC.

15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License

Lineage flow chart for mammalian embryogenetic development.

YOUNG AND BLACK

Fig. 1.
76

ADULT STEM CELLS
matogonia (male), undergo proliferation and subsequent
migration through various regions within the developing
conceptus, eventually reaching the gonadal ridges. Prolif-
eration, lineage commitment, and segregation of cells
within the inner cell mass lead to formation of two pluri-
potent layers, the epiblast and hypoblast. The epiblast
and hypoblast develop further by proliferation, lineage
commitment, and segregation into the primary pluripo-
tent germ layers: ectoderm, mesoderm, and endoderm
(Fig. 1). The ectodermal germ layer continues its develop-
ment by proliferation, lineage commitment, and segrega-
tion to form the surface ectoderm and neural ectoderm.
The neural ectoderm segregates into neural tube and neu-
ral crest. Similarly, the mesodermal germ layer continues
its development by proliferation, lineage commitment,
and segregation to form the somitic mesoderm, interme-
diate mesoderm, lateral plate somatic mesoderm, and lat-
eral plate splanchnic mesoderm. The endodermal germ
layer also continues its development by proliferation, lin-
eage commitment, and segregation. The cells of each of
these germ layer lineages undergo further rounds of pro-
liferation and segregation through lineage commitment to
form multipotent, tripotent, bipotent, and unipotent pro-
genitor cell lineages that finally form the differentiated
cells, tissues, and organs that are the end products of
these pathways of differentiation (Fig. 1).
Although a majority of the cells progress through this
preprogrammed developmental sequence to form an indi-
vidual, a few cells leave this developmental continuum to
become reserve precursor cells. These reserve precursor
cells are involved in the continual maintenance and repair
of tissues and organs throughout the life span of the
individual. It has long been assumed (and reiterated con-
tinually in review articles, most recently those concerning
embryonic stem cells) that the particular cells leaving the
developmental continuum to become reserve precursor
cells are limited solely to lineage-committed progenitor
cells specific for particular tissues. This assumption is
based in part on the previous identification of tissue-spe-
cific lineage-committed progenitor cells. Mauro (1961)
identified the myosatellite myoblast, a precursor cell for
the maintenance and repair of adult skeletal muscle. This
quiescent myogenic precursor cell is located between the
sarcolemma of the skeletal muscle cell and its adjoining
basement membrane. Chondrogenic precursor cells for the
maintenance and repair of cartilage were identified in the
perichondrial connective tissue surrounding cartilage
(Cruess, 1982). Osteogenic precursor cells for the mainte-
nance and repair of bone were identified in the outer
periosteal connective tissues covering bone (Cruess, 1982),
along the inner endosteal surface (Cruess, 1982), and
within the stroma of adjacent bone marrow (Owen and
Friedenstein, 1988). Adipogenic precursor cells for the
maintenance and generation of adipose tissue have been
identified within adult adipose tissue (Aihaud et al.,
1992). More recently, tissue-specific precursor cells have
been identified for other tissue types. Neurogenic precur-
sor cells have been located within tissues of the central
nervous system (Gage et al., 1995); epidermal precursor
cells have been identified within the interfollicular epider-
mis and bulge region of the hair follicle (Janes et al.,
2002); and basal epithelial cells have been identified as
the precursor cells for the gastrointestinal mucosa
(Bjerknes and Cheng, 2002). Based on these and other
reports, it has been assumed that each tissue and/or organ
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
77
within the postnatal individual contains only its own
unique contingent of precursor cells specific for that tis-
sue. It is further assumed that such lineage-committed
progenitor cells function solely in the maintenance and
repair of the tissue in which they reside (review, Donovan
and Gearhart, 2001; Forbes et al., 2002; Poulsom et al.,
2002; Tsai et al., 2002).
Young and his colleagues have been characterizing the
identities of precursor cells within the tissues of animals
of all ages, including humans. These studies began with
initial observations on the origins of the blastema cells
during limb regeneration in terrestrial amphibians
(Young, 1977a, 1977b, 1977c, 1983; Young et al., 1983a,
1983b, 1983c, 1983d, 1985, 1989a, 1989b). The studies
have progressed through precursor cell identification in
the connective tissues of avians (Bowerman et al., 1991;
Young et al., 1991, 1992a, 1992b, 1992c, 1993, 1995,
1998a; Lucas et al., 1994a; Dixson et al., 1996; Young and
Lucas, 1998; Young, 2000), mammals (Shoptaw et al.,
1991; Lucas et al., 1992, 1993, 1994a, 1994b, 1995; Pate et
al., 1993; Grande et al., 1995; Rogers et al., 1995; Ware-
jcka et al., 1996; Young and Lucas, 1998; Young et al.,
1998b, 2001a, 2004a, 2004b; Romero-Ramos et al., 2002;
Young, 2004), and humans (Young et al., 1999, 2001b,
Young, 2004). These studies have utilized numerous tech-
niques, including histology, energy-dispersive spectral
analysis coupled with scanning electron microscopy, gly-
coconjugate histochemistry, immunocytochemistry, quali-
tative and quantitative enzyme-linked immunoculture as-
says, differential isolation and plating, differential
cryopreservation, differential centrifugation, repetitive
limiting serial dilution clonogenic analysis, cluster of dif-
ferentiation (CD) marker analysis, fluorescent-activated
cell sorting, magnetic bead cell sorting, and molecular
analyses for telomerase activity and gene expression.
These techniques have been employed in studies of prolif-
eration, lineage commitment, lineage progression, and lin-
eage inhibition induced by bioactive factors. These studies
have led to the discovery that two general categories of
reserve precursor cells exist within the body and are in-
volved in the maintenance and repair of tissues in adults:
lineage-committed progenitor cells and lineage-uncommit-
ted pluripotent stem cells.
Lineage-committed progenitor stem cells display cer-
tain characteristics in common (Tables 1 and 2) (Young et
al., 1991, 1992a, 1992b, 1993, 1995, 1998a, 1998b, 1999,
2001a, 2001b, 2004a; Rogers et al., 1995; Young and Lu-
cas, 1998; Young, 2000, 2004). Progenitor cells may be
committed to one or more specific tissue lineages. Progen-
itor cells may be unipotent, forming a single cell type;
bipotent, forming two cell types; tripotent, forming three
cell types; or multipotent, forming four or more discrete
cell types. Each progenitor cell for a particular tissue
lineage has a unique profile of cell surface cluster of dif-
ferentiation (CD) markers. Progenitor cells conform to
Hayflick’s limit of 50 –70 population doublings (Hayflick,
1965) before they undergo programmed senescence and
cellular death. Progenitor cells are unresponsive to lin-
eage induction agents that have actions outside their re-
spective tissue lineage(s). For example, unipotent myo-
blast progenitor cells are unresponsive to bone
morphogenetic protein-2 (BMP-2), which induces cartilage
and bone development. Progenitor cells are responsive to
proliferation agents such as platelet-derived growth fac-
tors (i.e., PDGF-BB, PDGF-AA, PDGF-AB). Progenitor
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
78 YOUNG AND BLACK

TABLE 1. Postnatal precursor cells

Proliferation
Name Species Characteristics potential Differentiation potential References
Adult neuronal Rat Dependent on FGF-2 for survival, 1 year Astroglial, oligodendral Gage et al.,
progenitor cells Nestin⫹, A2B5⫹, O4⫹, GAD⫹, through 1995
NSE⫹, MAP2⫹, MAP5⫹, multiple
neuN⫺, tau⫺ passages
Adult liver- Mouse Ly5.1⫹, c-Kit⫹, Sca-1⫹, linlow/⫺ Not stated in lethally irradiated Taniguchi et
derived mice, replaced all al., 1996
hematopoietic blood cells (B cells, T
stem cells cells, granulocytes,
macrophages,
erythrocytes)
Adipo-fibroblasts Sheep Isolated by Percoll gradient Less than 2 Mature adipocytes Vierck et al.,
centrifugation weeks 1996
Mesenchymal Rat Selectively plated, released, ? 4–5 weeks Skeletal myotubes, Warejcka et
stem cells filtered, frozen and relplated adipocytes, endothelial al., 1996
cells, chondrocytes,
osteoblasts, smooth
muscle cells
Pancreatic- Mouse Dispersed islets grown to Not stated Pancreatic islets Cornelius et
derived “ . . . round cells budding containing ␣-cells, ␤- al., 1997
pluripotent upward from the epithelial cells, and ␦-cells
islet-producing monolayers . . . ”
stem cells
Neural stem cells Mouse Transgenic for lacZ, clonally Continuously B-lymphocytes, T- Bjornson et
derived (2H1), react strongly to expanded lymphocytes, myeloid al., 1999
X-Gal for cells
extended
periods of
time
SVZ astrocytes Mouse GFAP⫹, S100?⫹, vimentin⫹, Not stated TuJ1⫹, type A cells, type Doetsch et al.,
(neural stem labeled with 3H, DAP, GFP C cells, neurospheres, 1999
cells) neuroblasts
Multipotent stem Mouse Methylcellulose clonal spheres; 18–21 Form neuronal, Gritti et al.,
cell from FGFR1 & EGFR1 passages, astroglial, and 1999
subventricular ⱕ12 hour oligodendroglial cells
forebrain doubling
region time
Bone Marrow Mouse WT dystrophin gene ⫹ Not stated ⫹ Dystrophin fibers in Gussoni et al.,
Cells recipients 1999
Hematopoietic Mouse Sca-1⫹, c-Kit⫹, CD43⫹, CD45⫹, Not stated Dystrophin fibers in Gussoni et al.,
stem cells lin⫺ (B220⫺, Mac-1⫺, Gr1⫺, recipients 1999
CD4⫺, CD5⫺, CD8⫺)
Muscle side Mouse Sca-1⫹, lin⫺, c-Kit⫺, CD45⫺, Not stated myoblasts, fibroblasts, Gussoni et al.,
population CD43⫺ CD43⫹, CD45⫹, 1999
cells dystrophin ⫹
myofibers
Muscle main Mouse lin⫹ (CD11⫹, Gr1⫹, CD5⫹) Not stated Desmin ⫹ Gussoni et al.,
population 1999
cells
Ependymal- Rat, Notch-1, Decreased Olfactory bulb neurons, Johansson et
derived neural Mouse with neurons, astrocytes, al., 1999
stem cell increasing oligodendrocytes,
time after
injury
Marrow Stromal Mouse CD11b⫺, CD45⫺, BrdUrd labeled Not stated GFAP⫹, ORO⫹, Kopen et al.,
Cells BrdUrd⫹, 1999
Neurofilament⫹, Type
I & II collagen⫹,
Alizarin Red S⫹
Bone Marrow Rat Male, DPPIV⫹ Not stated Male DPPIV⫹ Petersen et
Stem Cells hepatocytes cells (also al., 1999
H4⫹, C-CAM⫹) found
in female DPPIV-
recipent
Mesenchymal Human SH2⫹, SH3⫹, CD29⫹, CD44⫹, Proliferate adipocytes, chondrocytes, Pittenger et
stem cells CD71⫹, CD90⫹, CD106⫹, extensively osteocytes al., 1999
CD120a⫹, CD124⫹, CD14⫺,
CD34⫺, CD45⫺, can be
subcloned at 4 cells per well
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
ADULT STEM CELLS 79
TABLE 1. Postnatal precursor cells (continued)
Proliferation
Name Species Characteristics Potential Differentiation Potential References
Mesenchymal Rat top 2/3 of Percoll gradient Not stated Muscle, tendon, Tomita et al.,
stem cells cartilage, bone, fat, 1999
cardiomyocytes
Pluripotent Human Sorted, CD10⫹, CD13⫹, CD34⫹, ⱕ 400 Mesodermal only: Muscle Young et al.,
Mesenchymal CD56⫹, CD90⫹, MHC-I⫹ population (3), adipose (2), 1999,
Stem Cells doublings Cartilage (5), bone (2), 2001b,
Connective tissue (4), 2004a;
Endothelial cells, Young,
Hematopoietic cells 2000, 2004
Skeletal muscle Mouse CD34⫹, Myf-5⫹, M-Cadherin⫹ Not stated Skeletal muscle Beauchamp et
satellite cells al., 2000
Neural stem cells Mouse b-Gal⫹, express neomycin Not stated Integrate into developing Clarke et al.,
resistance gene, Desmin⫹, chick embryos and 2000
Myosin heavy chain⫹, clonal contribute to all germ
culture layers
Bone marrow- Human FLK-1⫹, TRK⫹, TransferrinR⫹, “Rapidly Osteogenesis, Colter et al.,
derived: small Annexin-II⫹, CD44⫹, CD49e⫹, dividing adipogenesis, 2000, 2001
agranular CD59⫹, cells” chondrogenesis
recycling cells
(RS-1)
Bone marrow- Human FLK-1⫾, TRK⫾, CD49e⫹ “Rapidly Osteogenesis, Colter et al.,
derived: small dividing adipogenesis, 2000, 2001
gran. recycling cells” chondrogenesis
cells (RS-2)
Bone marrow- Human FLK-1⫺, TRK⫺, TransferrinR⫺, “Rapidly Osteogenesis, Colter et al.,
derived: large Annexin-II⫺, CD10⫹, CD147⫹, dividing adipogenesis, 2000, 2001
and more CD44⫹, CD49e⫹, CD59⫹, cells” chondrogenesis
mature cells CD81⫹, CD90⫹
(mMSCs)
Neural stem cells Mice Clonally derived, do not express 7–8 months MyHC⫹, MyoD⫹, Galli et al.,
and any myogenic markers, Myosin⫹, Laminin⫹ 2000
Human MLC3F/nlacZ⫺,
Muscle-derived Mouse Sca-1⫹, desmin⫹, CD34⫹, Bcl- 25 passages Muscle regeneration, Lee et al.,
stem cell clone, 2⫹, Flk-1⫹, c-met⫹, MNF⫹ osteoblasts, improve 2000
MC13 bone healing
Bone marrow- Human Clones - 3 types: osteogenic, 22–23 Bone; bone and cartilage; Muraglia et
derived osteo-chondrogenic; osteo- population and bone, cartilage, al.,2000
mesenchymal chondro-adipogenic doublings and adipocytes
progenitor cells
Islet-producing Mouse Insulin I⫹, Insulin II⫹, Insulin 3 years ⫹ Islet like structures that Ramiya et al.,
stem cells receptor⫹, hepatocyte growth secrete insulin 2000
factor⫹, C-MET⫹, glucagons⫹,
somatostatin⫹, glucose
transporter-2 receptor⫹,
glutamic acid decarboxylase-
67⫹, insulin-like growth factors
I & II⫹
Bone marrow Rat CD44⫹, CD71⫹, CD90⫹ ⬎ 20 Neuron-specific enolase Woodbury et
stromal cells passages (NSE⫹), NeuN⫹, al., 2000
nestin⫹, trkA⫹,
neurofilament-M⫹,
tau⫹
Bone marrow Human Not stated Passage 2 NSE⫹, NF-M⫹, growth Woodbury et
stromal cells cones, terminal bulbs al., 2000
Blood-derived Human Buffy coat isola: CD105⫹, SDF- Not stated Fibroblasts, adipocytes, Zvaifler et al.,
Mesenchymal 1⫹ osteoblasts/bone, 2000
precursor cells osteoclasts
Bone marrow- Mouse CD34⫺/low, c-Kit⫹, Sca-1⫹ Not stated Cardiac myocytes and Jackson et al.,
derived side endothelial cells 2001
population
cells
Muscle Precursor Mouse Sca-1⫹, CD34low/⫺, desmin⫹ Not stated dystrophin expressing Jankowski et
cells myofibers al., 2001
Bone marrow- Rat CD90⫹ 4,500 cells Osteogenesis Javazon et al.,
derived: small per cell 2001
recycling stem plated
cells (refed)
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
80 YOUNG AND BLACK

TABLE 1. Postnatal precursor cells (continued)

Proliferation
Name Species Characteristics Potential Differentiation Potential References
Bone marrow- Rat CD90⫹, CD59⫹ “Rapidly CFU formation, Javazon et al.,
derived: dividing osteogenic 2001
mature MSCs cells” differentiation
Marrow derived Mouse Fr25lin⫺, PKH26 labeled Not stated Incorporated onto bone Krause et al.,
stem cells marrow, stomach, 2001
esophagus, small
intestine, large
intestine, liver, lungs,
and skin
Circulating Mouse Blood via cardiac puncture, CFU Not stated Osteogenesis, Kuznetsov et
skeletal stem per 106 cells - 0.93 Adipogenesis al., 2001
cells
Circulating Rabbit Blood via cardiac puncture, CFU Not stated Osteogenesis, Kuznetsov et
skeletal stem per 106 cells - 0.18 Adipogenesis al., 2001
cells
Circulating Guinea Blood via cardiac puncture, CFU Not stated Osteogenesis, Kuznetsov et
skeletal stem Pig per 106 cells - 2.7 Adipogenesis al., 2001
cells
Circulating Human Whole blood & buffy coat conc., Not stated Osteogenesis, Kuznetsov et
skeletal stem CFU per 106 cells - rare, Adipogenesis al., 2001
cells
Neural Human Not stated 30–70⫹ cell neurons, astrocytes Palmer et al.,
progenitor cells doublings 2001
before
senescence
Multipotent Human long telomeres, CD45⫺, 70⫹ cell mesenchymal cell types, Reyes and
Adult glycophorin-A-, can be doublings cells of Verfaillie,
Progenitor subcloned at 10 cells per well neuroectodermal 2001
Cells lineage
Mesodermal Human CD45⫺, glycophorin-A⫺, CD10⫺, more than all mesodermal Reyes et al.,
progenitor cells CD31⫺, CD34⫺, CD36⫺, 50 cell phenotypes, 2001
CD38⫺, CD50⫺, CD62e⫺, doublings osteoblasts,
CD106⫺, CD117⫺, H1P12⫺, chondroblasts,
Fibroblast⫺, HLA-DR⫺, class 1 adipocytes, cells will
HLA⫺, Tie⫺, Tek⫺, b2- support hematopoiesis,
microglobulin low, CD44low, skeletal muscle,
CDw90low, KDRlow, Fltlow, endothelial cells
CD13⫹, CD49b⫹, average
telomere length of 11–15 kb,
normal karyotype
Human Human Nucleated Not stated Bone, cartilage Turgeman et
Mesenchymal al., 2001
Stem Cells
Spinal cord Rat Spinal cord gray matter isolate 24–30 hour Neurons (160 kDa-NF⫹) Vacanti et al.,
progenitor cells doubling and glia (GFAP⫹) 2001a
time, 4
weeks in
culture
Spore-like cells Rat 5 ␮m, pericellular mucopolysac- 12–36 hour “ . . . cells appear to Vacanti et al.,
charides and glycolipids doubling differentiate into cells 2001b
time specific to organ from
which they were
isolated, . . . ”
Pluripotent Rat Cloned, can be subcloned at 1 cell ⬎ 400 Mesodermal only: Muscle Young et al.,
Mesenchymal per well, Telomerase ⫹ population (3), adipose (2), 2001a,
Stem Cells doublings Cartilage (5), bone (2) 2004a;
Connective tissue (4), Young,
Endothelial cells 2000, 2004
Pluripotent Human CD10⫹, CD13⫹, CD34⫹, CD56⫹, ⬎ 400 Mesodermal only: Muscle Young et al.,
Mesenchymal CD90⫹, MHC-I⫹ population (3), adipose (2), 2001b,
Stem Cells doublings Cartilage (5), bone (2) 2004a;
Connective tissue (4), Young,
Endothelial cells, 2000, 2004
Hematopoietic cells
Adipose-derived Human Lipoaspirates: AS02⫹ (fibroblasts ⬍ 25 Adipogenic, Zuk et al.,
multi-lineage & mesenchymal cells); population chondrogenic, 2001
cells vimentin⫹ doublings myogenic, and
osteogenic cells
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
ADULT STEM CELLS 81
TABLE 1. Postnatal precursor cells (continued)
Proliferation
Name Species Characteristics Potential Differentiation Potential References
Nestin-positive Rat Nestin-positive, CK19-negative Longevity in Liver cells, pancreatic Zulewski et
islet-derived culture ⬎ acinar cells, ductal al., 2001
multipotential 8 months cells, ␣-cells, ␤-cells
stem cells
(NIP)
Nestin-positive Human Nestin-positive CK19-negative Longevity in Liver cells, pancreatic Zulewski et
islet-derived culture ⬎ acinar cells, ductal al., 20010
multipotential 8 months cells, ␣-cells, ␤-cells
stem cells
(NIP)
Multipotential Mice Induced somatic mutated cells Long lived, Form parietal, zymogen, Bjerknes and
stem cells 48⫹ enteroendocrine, and Cheng, 2002
weeks mucus cell types
Muscle-derived Mouse Sca-1⫹, slowly adhering, Not stated Muscle regeneration, Deasy et al.,
stem cells desmin⫹, osteoblasts, improve 2002
bone healing,
reconstitute
hematopoietic system
Human Stem Human CD45⫺, CD31⫺ Not stated CD90⫹, CD105⫹, Dell’Agnola et
Cells CD106⫹, CD133⫺, al., 2002
CD135⫺, HER2/neu⫹,
P1H12⫹, c-kit⫹,
CXCR4⫹,
Multipotent Mouse CD44⫺, CD45⫺, MHC-I⫺, MHC- 75 CD31⫹, Tau⫹, HNF-1⫺, Jiang et al.,
Adult II⫺, c-kit⫺, Ter1119⫺, CD13⫹, population GFAP⫹, CK18⫹, 2002
Progenitor Flk1dim, Oct4⫹, Rex1⫹, CD31⫺, doublings Flk1⫹, NF-200⫹,
Cells CD62E⫺, Tek⫺, vWF⫺ NSE⫹, vWF⫹,
Albumin⫹:
Endothelial,
astrocytes, neurons,
epithelioid
Cementum- Mouse Low levels or no alkaline Not stated Bone-like tissue with Krebsbach
derived cells phosphatase activity osteocytes/cemento- and Robey,
cyte-like cells 2002
Cementum- Human Low levels or no alkaline Not stated Bone-like tissue with Krebsbach
derived cells phosphatase activity osteocytes/cemento- and Robey,
cyte-like cells 2002
Dental pulp stem Human Collagen type XVIII ␣1, IGF-2, Not stated Dentin lined with Krebsbach
cells discordin domain tyrosine odontoblasts-like cells, and Robey,
kinease-2, cyclin-dependent dental pulp-like tissue 2002
kinase6
Neural Crest Rat p75 ⫹ Self- neurons, glia, Kruger et al.,
Stem Cells Renewal myofibroblasts 2002
declined
with
increasing
age
Bone marrow Mouse Unfractionated bone marrow Not stated Diploid, desmin, Myf-5, LaBarge and
derived cell isolate cMet-R, ␣7-integren, Blau, 2002
Satellite cell to muscle
repair
Progenitor Cells Rat Cells from low-buoyancy fraction Not stated NeuN⫹, Rip ⫹/⫺, Lie et al.,
of Percoll, BrdU⫹, GFP⫹, NG2⫹, APC⫹, 2002
NG2⫹ S100b⫹, GFAP⫹,
A2B5⫹, BrdU⫹,
Ox42⫹/⫺, Nestin⫹
TH⫹, b-Tubulin III⫹/
⫺
Muscle-derived Mouse Sca-1⫹, CD45⫹, CD34⫺, c-kit⫺ Not stated Hematopoietic cells: McKinney-
hematopoietic Gr.1⫹, Mac-1⫹, Freeman et
stem cells Thy1⫹, B220⫹ al., 2002
Muscle-derived Mouse CD45⫺, Sca-1⫹ & CD45⫺, Sca- Not stated Fibroblastic-like cells McKinney-
myogenic stem 1⫺ and twitching skeletal Freeman et
cells muscle myofibers al., 2002
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
82 YOUNG AND BLACK

TABLE 1. Postnatal precursor cells (continued)

Proliferation
Name Species Characteristics Potential Differentiation Potential References
Neural Stem Mouse Nestin⫹, EGFP⫹, b-Tubulin Self- Neurospheres Murayama et
Cells III⫹, GFAP⫺, O4⫹ Renewal al., 2002
of sorted
neurospher
e-
initiating
cells
Neural Stem Cell Mouse c-Kit⫹, Sca1⫺, CD45⫺, Notch Not stated Unknown Murayama et
Side 1⫹, Nestin⫺, EGFP⫺ al., 2002
Population
Bone Marrow- Mouse CD3⫺, CD11⫺, CD45⫺, Ter- Not stated CD31⫹, c-kit⫹, Sca1⫹, Otani et al.,
derived stem 199⫺, Ly-6G⫺ Flk1⫹ 2002
cells
Bone Marrow Human ␣-LNGFR ⫹, CD45⫺, a- High CD45⫺, TE7⫹, CD14⫹, Quirici et al.,
Mesenchymal glycophorin-A⫺, SH2⫹, proliferative CD34⫺, CD133⫺, 2002
Stem Cells ␣- CD34⫹, CD133⫹ capacity ORO⫹, Alizarin Red S
LNGFR⫹ ⫹
Bone Marrow Human ␣-LNGFR⫺, SH2⫺, CD45⫺, a- High CD45⫺, TE7⫹, CD14⫹, Quirici et al.,
Mesenchymal glycophorin-A⫺ proliferative Alizarin Red S ⫹ 2002
Stem Cells ␣- capacity
LNGFR⫺
Pluripotent Stem Rat Vimentin⫹, Pax6⫹, CD34 varied, Not stated Nestin⫹, Pax6⫹, Oct4⫹, Romero-
Cells CD45⫺, CD90⫹, Oct4 low, MBP⫹, NG2⫹, Ramos et
Myo-D⫺, myogenin⫺ NF145⫹, CNPase⫹, al., 2002
Tuj1⫹, NF68⫹,
GFAP⫹, MOSP⫹,
Tau⫹, NeuN⫺
Smooth Muscle Human CD34⫹, CD31⫺, Tie2⫺, Flk1⫹, Not stated Smooth muscle cells Simper et al.,
Outgrowth Flt1⫹, VE-Cadherin⫺, vWF⫺, 2002
Cells VEGF1⫹,2⫹, Integrin␣5␤1⫹,
␣SMA⫹, MHC⫹, Calponin⫹
Endothelial Human CD31⫹, vWF⫹, VE-Cadherin⫹, Not stated Endothelial cells Simper et al.,
Outgrowth Flt1⫹, Flk1⫹, ␣SMA⫺, Tie-2⫹ 2002
Cells
Bone marrow Rat Ring cloning; express germline, 15 to ⬎20 Induced neurons: Tau⫹, Woodbury et
stromal cells ectodermal, mesodermal, & passages TOAD-64⫹, b-Tubulin al., 2002
endodermal genes prior to III⫹, synaptophysis⫹,
differentiation ChAT⫹, TH⫹,
GFAP⫹, NF-M⫹,
vK⫹, NeuroD⫺
Adult hepatic Rat Two-step collagenase-perfusion, Long term Induced pancreatic Yang et al.,
oval stem cells cell sorting with Thy-1.1: AFP, culture for endocrine hormone 2002
albumin, ␥-glutamyl- ⬎ 1 year producing cells:
transpeptidase CK19, OV6 by serial insulin-1, insulin-2,
1:2 splits glucagon, somatostatin
Neural stem cells Human Not stated 60 cell Not stated Johe, 2003
doublings
Muscle SP cells Mouse SP⫹ (side population⫹), CD34⫺, Not stated Endothelium Majka et al.,
c-met⫹, CD45⫹, Sca-1⫹, PE- 2003
CAM⫹, Tie-2⫹
Muscle non-SP Mouse SP⫺, c-met⫹, CD45⫹, Sca-1⫹, PE- Not Stated Smooth muscle Majka et al.,
cells CAMlow, Tie-2low, CD34low, SM- 2003
␣-actin⫹, PDGFR␤⫹, Calponin-
mRNA
Hepatic stellate Mouse Anti-␣-smooth muscle actin⫹, ? 7 days When sitmulated with Oben et al.,
cells GFAP⫹ serum and PDGF the 2003
cells increased
proliferation and
increased expression of
collagen 1␣2
Adipose tissue- Human From low density region of Ficoll- 2 weeks Adipogenic, osteogenic, Winter et al.,
derived Paque Plus density gradient chondrogenic 2003
Stromal cells
Pluripotent Rat Cloned, can be subcloned at 1 cell ⬎ 400 Ectoderm: 7⫹ cell types; Young, 2004;
Epiblastic-Like per well, Oct-4⫹, Telomerase⫹ population Mesoderm: 18⫹ cell Young et
Stem Cells doublings types; Endoderm: 11⫹ al., 2004b
cell types
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
ADULT STEM CELLS 83
TABLE 1. Postnatal precursor cells (continued)
Proliferation
Name Species Characteristics Potential Differentiation Potential References
Pluripotent Human Sorted, CD10⫹, CD66e⫹ SSEA- ⬎ 400 Ectoderm: 7⫹ cell types; Young, 2004;
Epiblastic-Like 1⫹, SSEA-3⫹, SSEA-4⫹, population Mesoderm: 18⫹ cell Young et
Stem Cells CEA⫹, HCEA⫹, CEA-CAM1⫹ doublings types; Endoderm: 11⫹ al., 2004a
cell types
Adult pluripotent Human CD14, CD34, CD45 Not stated Macrophages, T- Zhao et al.,
stem cells lymphocytes, epithelial 2003
cells, endothelial cells,
neuronal cells, liver
cells

TABLE 2. Precursor cell characteristics

Progenitor Cells PPGLSCs1 PPELSCs2
Serum-Free Defined Medium3 Quiescence Quiescence Quiescence
Commitment Lineage-Specific Germ Layer-Specific Uncommitted
Life Span4 Hayflick’s Limit5 Extended6 Extended
Telomerase Absent Present Present
Cell Growth at Confluence Contact Inhibited Contact Inhibited Non-Contact Inhibited
Proliferation7 Responsive Responsive Responsive
Induction8 Unresponsive Responsive, Germ Layer-specific Responsive, unlimited
Progression9 Responsive Unresponsive Unresponsive
Inhibitory10 Responsive Responsive Responsive
Antibodies11 Cell-Specific Germ Layer-Specific Embryonic
CD Markers12 Cell-Specific CD10, CD13, CD34, CD56, CD90, CD10, CD66e
MHC1 (PPMSCs only)
Cells Formed Lineage-Specific Germ Layer-Specific Ectoderm, Mesoderm,
Endoderm
1
PPGLSCs, pluripotent germ layer stem cells, i.e., pluripotent ectodermal stem cells (PPEctoSCs), pluripotent mesodermal
stem cells (PPMSCs), pluripotent endodermal stem cells (PPEndoSCs).
2
PPELSCs, pluripotent epiblastic-like stem cells.
3
Serum-Free Defined Medium, medium in the absence of proliferation factors, induction factors, progression factors, and
inhibitory factors (i.e., leukemia inhibitory factor, fibroblast feeder layer, anti-differentiation factor, scar inhibitory factor, etc.)
or their activities.
4
Life Span, population-doubling number.
5
Hayflick’s Limit, up to 50-70 population doublings before cell senescence and cellular death (Hayflick, 1965).
6
Extended, population doublings greater than Hayflick’s limit of 70. In this instance both PPMSC clone and PPELSC clone
were tested to 400 population doublings. Both clones retained their respective pluripotent potentials throughout maximum
population doublings examined (Young, 2004).
7
Proliferation, initiation of cell proliferation in response to proliferation agents such as platelet-derived growth factors
(PDGF)-AA, PDGF-BB, PDGF-AB, etc.
8
Induction, commitment to a specific germ layer or tissue/cell lineage in response to a specific (i.e., bone morphogenetic
protein-2, skeletal muscle morphogenetic protein, etc.) or a general (dexamethasone, etc.) lineage-tissue-cell inductive agent.
9
Progression, acceleration of expression of specific phenotypic marker(s) in response to progression agents such as insulin,
insulin-like growth factor (IGF)-I, IGF-II, etc.
10
Inhibitory, inhibition of commitment to and/or expression of specific phenotypic marker(s) in response to a general (i.e.,
leukemia inhibitory factor, LIF; ESGRO, murine LIF; fibroblast feeder layer; anti-differentiation factor, ADF; etc.) or a specific
(scar inhibitory factor, SIF; etc.) inhibitory agent.
11
Antibodies, antibodies to phenotypic expression markers, can be cell specific (i.e., OP137, F5D, MF-20, MY-32, ALD-58, and
A4.74 for skeletal muscle), germ layer specific (i.e., FORSE-1, Rat-401, HNES, and MAB353 for neural ectoderm), or embryonic
(i.e., SSEA-1, SSEA-3, SSEAS-4, CEA, HCEA, CD66e, and CEA-CAM-1 for embryonic stem cells) (see Table 3 for further
examples).
12
CD Markers, cluster of differentiation makers.

cells exhibit contact inhibition at confluence. Progenitor
cells are responsive to progression agents (i.e., insulin,
insulin-like growth factor-I (IGF-I), insulin-like growth
factor-II (IGF-II)) that accelerate the time frame of expres-
sion for tissue-specific markers of phenotypic differentia-
tion. For example, skeletal muscle myoblasts grown in the
presence of IGF-I will demonstrate myogenic phenotypic-
specific expression markers characteristic of skeletal mus-
cle (i.e., sarcomeric myosin, myosin heavy chain, myosin
fast chain, etc.) in about half the time taken by myoblasts
grown in the absence of IGF-I. Progenitor cells remain
quiescent in a serum-free environment lacking progres-
sion agents, proliferation agents, and inhibitory factors
(i.e., leukemia inhibitory factor, murine leukemia inhibi-
tory factor (ESGRO), a fibroblast feeder layer, antidiffer-
entiation factor, etc.).
Examples of lineage-committed progenitor cells include
the unipotent myosatellite myoblasts of muscle (Mauro,
1961; Campion, 1984; Grounds et al., 1992; Young et al.,
1993, 1995, 1998b); the unipotent adipoblast cells of adi-

84 YOUNG AND BLACK
pose tissue (Ailhaud et al., 1992; Young et al., 1993, 1995);
the unipotent chondrogenic cells and osteogenic cells of
the perichondrium and periosteum, respectively (Cruess,
1982; Young et al., 1993, 1995); the unipotent basal epi-
thelial cells of gastrointestinal mucosa (Bjerknes and
Cheng, 2002); and neuronal unipotent progenitor cells
(Gage et al., 1995). Bipotent progenitor cells include the
adipofibroblasts of adipose tissue (Vierck et al., 1996) and
the bipotent chondrogenic-osteogenic stem cells of marrow
(Owen, 1988; Owen and Friedenstein, 1988; Beresford,
1989; Caplan et al., 1997; Prockop, 1997; Young, 2000).
Tripotent progenitor cells include the chondrogenic-osteo-
genic-adipogenic stem cells of marrow (Pittenger et al.,
1999; Young, 2000). Multipotent hematopoietic cells of
marrow (McGuire, 1998; Palis and Segel, 1998; Ratajczak
et al., 1998) are an example of multipotent progenitor
cells. See Table 1 for additional examples of lineage-com-
mitted progenitor cells.
During the studies that led to the characterization of
lineage-committed progenitor cells (Young et al., 1991,
1992a, 1992b, 1993, 1995, 1998a, 1998b, 1999, 2001a,
2001b, 2004a; Rogers et al., 1995; Young and Lucas, 1998;
Young, 2000, 2004), we noted the presence of another
general category of precursor cells within the tissues, the
lineage-uncommitted pluripotent stem cells. Further ex-
periments revealed the presence of unique subcategories
of these pluripotent stem cells. To date we have been
concerned with the definitive characterization of two of
these subcategories of pluripotent stem cells. Both subcat-
egories of pluripotent stem cells share a few similarities
with progenitor cells, including quiescence in an environ-
ment that lacks serum, proliferation agents, inductive
agents, progression agents, and inhibitory agents or their
influences. They are also similar in their response to pro-
liferation agents. However, these two pluripotent stem
cell subcategories differ in many respects from lineage-
committed progenitor cells (Tables 1 and 2). They also
demonstrate similarities and differences with respect to
each other (Tables 1–3).
Both pluripotent stem cell subcategories are telomerase
positive (Fig. 2A). Both exhibit extended capabilities for
self-renewal without loss of differentiative capabilities, far
surpassing Hayflick’s limit of 50 –70 population doublings.
Both pluripotent stem cell subcategories are unresponsive
to progression factors such as insulin, IGF-I, or IGF-II
that normally accelerate phenotypic expression in lineage-
committed progenitor cells.
The first subcategory of lineage-uncommitted pluripo-
tent stem cells characterized by Young and colleagues
(Young and Lucas, 1998; Young et al., 1998a, 1998b, 1999,
2001a, 2001b, 2004a; Young, 2000, 2004) express the fol-
lowing cell surface profile: CD10⫹, CD13⫹, CD34⫹,
CD56⫹, CD90⫹, MHC-I⫹, CD1a–, CD2–, CD3–, CD4–,
CD5–, CD7–, CD8–, CD9–, CD11b–, CD11c–, CD14–,
CD15–, CD16–, CD18–, CD19–, CD20–, CD22–, CD23–,
CD24–, CD25–, CD31–, CD33–, CD36–, CD38–, CD41–,
CD42b–, CD45–, CD49d–, CD55–, CD57–, CD59–, CD61–,
CD62E–, CD65–, CD66e–, CD68–, CD69–, CD71–, CD79–,
CD83–, CD95–, CD105–, CD117–, CD123–, CD135–, CD166–,
Glycophorin-A–, HLA-DRII–, FMC-7–, Annexin-V–, and
LIN–. These precursor cells are 10 –20 ␮m in size as de-
termined by flow cytometric analysis of unfixed cells.
These precursor cells do not express embryonic stem cell
markers such as alkaline phosphatase, stage-specific em-
bryonic antigen-1 (SSEA-1), SSEA-3, SSEA-4, carcinoem-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
bryonic antigen, or carcinoembryonic antigen-cell adhe-
sion molecule-1 (Tables 1–3). They exist in a quiescent
state of stasis in serum-free defined medium lacking pro-
liferation factors, inductive factors, progression factors, or
inhibitory factors. In this state they display no cell prolif-
eration, no spontaneous differentiation, and no cell death
(Fig. 3A). These cells display contact inhibition at conflu-
ence, ceasing cell proliferation once a single layer of cells
covers the surface of a flask (Fig. 3B), even in the contin-
ued presence of proliferation agents such as PDGFs.
These precursor cells are responsive to both general and
specific inductive factors, but only with respect to cells of
the mesodermal lineage. They can be induced to form 18 or
more phenotypes limited to the mesodermal lineage (Ta-
ble 3, Fig. 3C–T) from a single cell, derived from either
repetitive limiting serial dilution clonogenic analysis or
cell sorting. These cells will only form progenitor cells
committed to the mesodermal lineage, being unresponsive
to general or specific inductive factors for cells of the
ectodermal or endodermal lineages. Once committed to a
particular mesodermal tissue lineage (i.e., myogenic,
chondrogenic, etc.), the newly induced lineage-specific pro-
genitor cells are unresponsive to inductive factors outside
that respective lineage. For example, newly induced pro-
genitor cells committed to the myogenic lineage are unre-
sponsive to BMP-2, a factor inducing commitment to the
bipotent chondrogenic-osteogenic lineage. Similarly,
newly induced progenitor cells committed to the chondro-
genic lineage are unresponsive to skeletal muscle morpho-
genetic protein (Sk-MMP), a factor inducing commitment
to the skeletal muscle myogenic lineage. However, these
newly induced progenitor cells respond to progression fac-
tors by accelerating the expression of their differentiated
phenotype. For example, myogenic progenitor cells ex-
press differentiated muscle markers in about two weeks in
the presence of IGF-I rather than the usual four weeks in
the absence of a progression agent. Similarly, in the pres-
ence of IGF-I, osteogenic progenitor cells express markers
for differentiated bone in about four weeks rather than six
weeks in the absence of a progression agent. These newly
induced progenitor cells assume Hayflick’s limit of 50 –70
population doublings before programmed senescence and
cellular death occur. We designated these lineage-uncom-
mitted precursor cells as pluripotent mesodermal stem
cells (PPMSCs) due to their extended capabilities for self-
renewal that far surpass Hayflick’s limit and their ability
to form any cell type belonging to the mesodermal lineage.
The second subcategory of lineage-uncommitted precur-
sor cells undergoing characterization (Young et al., 2004a;
Young, 2004) expresses the following cell surface profile:
CD10⫹, CD66e⫹, CD1a–, CD2–, CD3–, CD4–, CD5–, CD7–,
CD8–, CD9–, CD11b–, CD11c–, CD13–, CD14–, CD15–,
CD16–, CD18–, CD19–, CD20–, CD22–, CD23–, CD24–,
CD25–, CD31–, CD33–, CD34–, CD36–, CD38–, CD41–,
CD42b–, CD45–, CD49d–, CD55–, CD56–, CD57–, CD59–,
CD61–, CD62E–, CD65–, CD68–, CD69–, CD71–, CD79–,
CD83–, CD90–, CD95–, CD105–, CD117–, CD123–,
CD135–, CD166–, Glycophorin-A–, MHC-I–, HLA-DRII–,
FMC-7–, Annexin-V–, and LIN–. A clone with similar at-
tributes to the CD10⫹, CD66e⫹ human precursor cell sub-
category was derived from postnatal rats by limiting serial
dilution clonogenic analysis. Both the sorted adult human
cells and the cloned postnatal rat cells show similarities
and differences with respect to the PPMSCs characterized
previously. The similarities include the presence of telom-
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
ADULT STEM CELLS 85
TABLE 3. Identification of phenotypic expression markers in postnatal pluripotent stem cells
Phenotypic Markers PPELSCs1 PPEctoSCs2 PPMSCs3 PPEndoSCs4
Embryonic
Alkaline Phosphatase ⫹ ⫺ ⫺ ⫺
SSEA-15 ⫹ ⫺ ⫺ ⫺
SSEA-36 ⫹ ⫺ ⫺ ⫺
SSEA-47 ⫹ ⫺ ⫺ ⫺
CEA8 ⫹ ⫺ ⫺ ⫺
HCEA9 ⫹ ⫺ ⫺ ⫺
CD66e10 ⫹ ⫺ ⫺ ⫺
CEA-CAM111 ⫹ ⫺ ⫺ ⫺
Oct-412 ⫹ nd13 nd nd
Ectoderm
Neurogenic Progenitor Cells14 ⫹ ⫹ ⫺ ⫺
Neurons15 ⫹ ⫹ ⫺ ⫺
Ganglia16 ⫹ ⫹ ⫺ ⫺
Oligodendrocytes17 ⫹ ⫹ ⫺ ⫺
Astrocytes18 ⫹ ⫹ ⫺ ⫺
Radial Glial Cells19 ⫹ ⫹ ⫺ ⫺
Keratinocytes20 ⫹ ⫹ ⫺ ⫺
Mesoderm
Skeletal Muscle21 ⫹ ⫺ ⫹ ⫺
Smooth Muscle22 ⫹ ⫺ ⫹ ⫺
Cardiac Muscle23 ⫹ ⫺ ⫹ ⫺
White Fat24 ⫹ ⫺ ⫹ ⫺
Brown Fat25 ⫹ ⫺ ⫹ ⫺
Hyaline Cartilage26 ⫹ ⫺ ⫹ ⫺
Articular Cartilage27 ⫹ ⫺ ⫹ ⫺
Elastic Cartilage28 ⫹ ⫺ ⫹ ⫺
Growth Plate Cartilage29 ⫹ ⫺ ⫹ ⫺
Fibrocartilage30 ⫹ ⫺ ⫹ ⫺
Endochondral Bone31 ⫹ ⫺ ⫹ ⫺
Intramembranous Bone32 ⫹ ⫺ ⫹ ⫺
Tendon/Ligament33 ⫹ ⫺ ⫹ ⫺
Dermis34 ⫹ ⫺ ⫹ ⫺
Scar Tissue35 ⫹ ⫺ ⫹ ⫺
Endothelial Cells36 ⫹ ⫺ ⫹ ⫺
Hematopoietic Cells37 ⫹ ⫺ ⫹ ⫺
Endoderm
Endodermal Progenitor Cells38 ⫹ ⫺ ⫺ ⫹
GI Epithelium39 ⫹ ⫺ ⫺ ⫹
Liver Oval Cells40 ⫹ ⫺ ⫺ ⫹
Liver Hepatocytes41 ⫹ ⫺ ⫺ ⫹
Liver Biliary Cells42 ⫹ ⫺ ⫺ ⫹
Liver Canalicular Cells43 ⫹ ⫺ ⫺ ⫹
Pancreatic Progenitor Cells44 ⫹ ⫺ ⫺ ⫹
Pancreatic Ductal Cells45 ⫹ ⫺ ⫺ ⫹
Pancreatic ␤-Cells46 ⫹ ⫺ ⫺ ⫹
Pancreatic ␣-Cells47 ⫹ ⫺ ⫺ ⫹
Pancreatic ␦-Cells48 ⫹ ⫺ ⫺ ⫹
1
PPELSCs, pluripotent epiblastic-like stem cells (isolated and cloned) (Young et al., 2004a, 2004b; Young, 2004).
2
PPEctoSCs, pluripotent ectodermal stem cells (induced) (Romero-Ramos et al., 2002; Young, 2004; Young et al., 2004a).
3
PPMSCs, pluripotent mesodermal stem cells (isolated and cloned) (Young et al., 1999, 2001a,2001b; Young, 2000, 2004).
4
PPEndoSCs, pluripotent endodermal stem cells (induced) (Young, 2004; Young et al., 2004a).
Embryonic cells were identified as follows.
5
SSEA-1, stage-specific embryonic antigen-1, MC480, Developmental Studies Hybridoma Bank (DSHB), Iowa City, IA (Solter
and Knowles, 1978).
6
SSEA-3, stage-specific embryonic antigen-3, antibody MC631 (DSHB) (Damjanov et al., 1982).
7
SSEA-4, stage-specific embryonic antigen-4, antibody MC-813-70 (DHSB) (Lannagi et al., 1983).
8
CEA, carcinoembryonic antigen, (Hixson) (Estrera et al., 1999).
9
HCEA, human carcinoembryonic antigen (Sigma) (Young et al., 2004a).
10
CD66e, carcinoembryonic antigen (Vector) (Kishimoto et al., 1997).
11
CEA-CAM1, carcinoembryonic antigen-cell adhesion molecule (Hixson) (Estrera et al., 1999).
12
Oct-4, a gene directly involved in the capacity for self-renewal and totipotency of mammalian embryonic stem cells (Pesce and
Scholer, 2001; Young et al., 2004a).
13
nd, not as yet determined.
Ectodermal lineage cells were identified as follows.
14
Neurogenic Progenitor Cells, were identified using FORSE-1 (DSHB) for neural precursor cells (Tole et al., 1995; Tole and
Patterson, 1995), RAT-401 (DSHB) for nestin (Hockfield and McKay, 1985), HNES (Chemicon, Temecula, CA) for nestin
(Young et al., 2004a,b), and MAB353 (Chemicon) for nestin (Gritti et al., 1996).
 15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
86 YOUNG AND BLACK

erase activity (Fig. 2A); extended capabilities for self-re-
newal far surpassing Hayflick’s limit; quiescence and sta-
sis in serum-free defined medium lacking proliferation,
induction, and inhibitory factors; unresponsiveness to pro-
gression factors; responsiveness to inductive factors and
inhibitory factors; and assumption of progenitor cell sta-
tus once committed to a particular tissue lineage. Once
committed to a particular tissue lineage, the progenitor
cells derived from the CD10⫹, CD66e⫹ lineage exhibit
contact inhibition at confluence, lack of responsiveness to
induction factors outside their respective tissue lineage,
responsiveness to progression factors accelerating their
phenotypic expression, and conformance to Hayflick’s
limit of 50 –70 population doublings before programmed
senescence and cellular death occur.
However, distinct differences also exist between the plu-
ripotent mesenchymal stem cells and the cells exhibiting
CD10⫹, CD66e⫹ cell surface markers or their equivalent
clone. The adult-derived CD10⫹, CD66e⫹ precursor cells
are highly mobile and small (Fig. 4A and B), averaging
6 – 8 mm in size by flow cytometric analysis of unfixed
cells. They do not exhibit contact inhibition at confluence,
but rather form multiple confluent layers of cells in the
presence of proliferation agents (Fig. 4C and D). Although
derived from adults, these precursor cells express embry-
onic stem cell markers, i.e., alkaline phosphatase,

15
Neurons, were identified using 8A2 (DSHB) for neurons (Drazba et al., 1991), S-100 (Sigma) for neurons (Baudier et al.,
1986), T8660 (Sigma) for beta-tubulin III (Banerjee et al., 1988, 1990; Joshi and Cleveland, 1990), RT-97 (DSHB) for
neurofilaments (Wood and Anderton, 1981), N-200 (Sigma) for neurofilament-200 (Debus et al., 1983a; Franke, et al., 1991),
and SV2 (DSHB) for synaptic vesicles (Feany et al., 1992).
16
Ganglia, were identified using TuAg1 (Hixson) for ganglion cells (Faris et al., 1990; Hixson et al., 1990).
17
Oligodendrocytes, were identified using Rip (DSHB) for oligodendrocytes (Friedman et al., 1989) and CNPase (Sigma) for
oligodendrocytes and astroglia (Sprinkle et al., 1987; Sprinkle, 1989; Reynolds et al., 1989).
18
Astrocytes, were identified using CNPase (Sigma) for astroglia and oligodendrocytes (Sprinkle et al., 1987; Sprinkle, 1989;
Reynolds et al., 1989).
19
Radial Glial Cells, were identified using 40E-C (DSHB) for radial glial cells (Alvarez-Buylla et al., 1987).
20
Keratinocytes, were identified using VM-1 (DSHB) to keratinocyte cell surface protein (Morhenn, 2002).
Mesodermal lineage cells were identified as follows.
21
Skeletal Muscle, was identified as mononucleated myoblasts staining with OP137 (Calbiochem, San Diego, CA) for MyoD
(Thulasi et al., 1996), F5D (DSHB) for myogenin (Wright et al., 1991), and DEU-10 (Sigma) for desmin (Debus et al., 1983b),
and as multinucleated spontaneously contracting structures staining with MF-20 (DSHB) for sarcomeric myosin (Bader et al.,
1982), MY-32 (Sigma) for skeletal muscle fast myosin (Naumann and Pette, 1994), ALD-58 (DSHB) for myosin heavy chain
(Shafiq et al., 1984), and A4.74 (DSHB) for myosin fast chain (Webster et al., 1988).
22
Smooth Muscle, was identified as mononucleated cells staining with antibodies IA4 (Sigma) for smooth muscle alpha-actin
(Skalli et al., 1986) and Calp (Sigma) for calponin (Frid et al., 1992; Lazard et al., 1993).
23
Cardiac Muscle, was identified as binucleated cells co-staining with MF-20 (DSHB) ⫹ IA4 (Sigma) for sarcomeric myosin and
smooth muscle alpha actin (Eisenberg and Markwald, 1997; Eisenberg et al., 1997), MAB3252 (Chemicon) for cardiotin
(Schaart et al., 1997) and MAB1548 for cardiac muscle (Chemicon).
24
White Fat, also denoted as unilocular adipose tissue, was identified as a mononucleated cell with a peripherally-located
nucleus and containing a large central intracellular vacuole filled with refractile lipid and stained histochemically for
saturated neutral lipid using Oil Red-O (Sigma) and Sudan Black-B (Chroma-Gesellschaft, Roboz Surgical Co, Washington,
DC) (Young et al., 2001a).
25
Brown Fat, also denoted as multi-locular adipose tissue was identified as a mononucleated cell with a centrally-located
nucleus containing multiple small intracellular vacuoles filled with refractile lipid and stained histochemically for saturated
neutral lipid using Oil Red-O (Sigma) and Sudan Black-B (Chroma-Gesellschaft) (Young, 2000; Young et al., 2001b).
26 –30
Cartilage: structures thought to be cartilage nodules were tentatively identified as aggregates of rounded cells containing
pericellular matrix halos. Cartilage nodules were confirmed both by both histochemical and immunochemical staining.
Histochemically, cartilage nodules were visualized by staining the pericellular matrix halos for proteoglycans containing
glycosaminoglycan side chains with chondroitin sulfate and keratan sulfate moieties. This was accomplished using Alcian Blue
(Alcian Blue 8GS, Chroma-Gesellschaft), Safranin-O (Chroma-Gesellschaft) at pH 1.0, and Perfix/Alcec Blue. Verification of
glycosaminoglycans specific for cartilage was confirmed by loss of extracellular matrix staining following digestion of the
material with chondroitinase-AC (ICN Biomedicals, Cleveland, OH) and keratinase (ICN Biomedicals) (Young et al., 1989a,b,
2001a,b) prior to staining (negative staining control). Immunochemically, the chondrogenic phenotype was confirmed by initial
intracellular staining followed by subsequent staining of the pericellular and extracellular matrices with CIIC1 (DSHB) for
type-II collagen (Holmdahl et al., 1986), HC-II (ICN Biomedicals, Aurora, OH) for type-II collagen (Burgeson and Hollister,
1979; Kumagai et al., 1994), D1-9 (DSHB) for type-IX collagen (Ye et al., 1991), 9/30/8A4 (DSHB) for link protein (Caterson
et al., 1985), 12/21/1C6 (DSHB) for proteoglycan-hyaluronate binding region (Caterson, 2001), and 12C5 (DSHB) for versican
(Asher et al., 1995). Types of cartilage were segregated based on additional attributes.
26
Hyaline Cartilage, was identified by a perichondrial-like connective tissue surrounding the above stained cartilage nodule
and histochemical co-staining for type-I collagen (Young et al., 1989c).
27
Articular Cartilage, was identified as the above stained cartilage nodule without a perichondrial-like connective tissue
covering (Young et al., 1993).
28
Elastic Cartilage, was identified by nodular staining for elastin fibers and a perichondrial-like connective tissue surrounding
the above stained cartilage nodule and histochemical co-staining for type-I collagen (Young et al., 1989c).
29
Growth Plate Cartilage, was identified by nodular staining for cartilage phenotypic markers (see above) and co-staining for
calcium phosphate using the von Kossa procedure (Young et al., 1999, 2001a,b).
30
Fibrocartilage, was identified as three-dimensional nodules demonstrating extracellular histochemical staining for type-I
collagen (Young et al., 1989c) and co-staining for pericellular matrices rich in chondroitin sulfates A and C. The latter were
assessed by Alcian Blue pH 1.0 staining. Negative staining controls were digested prior to staining with chondroitinase-ABC
or chondroitinase-AC (Young et al., 1989a,b, 2001a,b).

ADULT STEM CELLS
SSEA-1, SSEA-3, SSEA-4, carcinoembryonic antigen-cell
adhesion molecule-1, carcinoembryonic antigens (Table 1,
Fig. 4E–L), and Oct-4 gene expression (Fig. 2B and C), in
a quiescent undifferentiated state. To date, these precur-
sor cells have formed over 36 distinct cell types derived
from all three primary germ layer lineages: 7 or more
ectodermal cell types (i.e., neuronal progenitor cells, neu-
rons, ganglia, astrocytes, oligodendrocytes, radial glial
cells, keratinocytes) (Table 3, Fig. 5A–S), 18 or more me-
sodermal cell types (i.e., skeletal muscle, cardiac muscle,
smooth muscle, white fat, brown fat, hyaline cartilage,
elastic cartilage, growth plate cartilage, articular carti-
lage, fibrocartilage, cortical bone, trabecular bone, loose
fibrous connective tissues, tendon, ligament, scar-connec-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
87
tive tissue, endothelial cells, hematopoietic cells) (Table 3,
Fig. 6A–P), and 11 or more endodermal cell types (i.e.,
endodermal progenitor cells, gastrointestinal epithelial
cells, pancreatic progenitor cells, insulin-secreting ␤-cells,
glucagon-secreting ␣-cells, somatostatin-secreting ␦-cells,
pancreatic ductal cells, liver oval cells, liver hepatocytes,
liver biliary cells, and liver canalicular cells) (Table 3, Fig.
7A–P). We propose that the second subcategory of lineage-
uncommitted precursor cells is a sequestered embryonic-
like stem cell existing within an adult. We have desig-
nated this population as a pluripotent epiblastic-like stem
cell, due to its capacity for extended self-renewal well past
Hayflick’s limit and its ability to form cell types from all
three primary germ layer lineages.

31
Endochondral Bone, was identified as the formation of a three-dimensional structure with progressional staining from one
displaying chondrogenic phenotypic markers, i.e., pericellular type-II collagen, type-IX collagen, chondroitin sulfate/keratan
sulfate glycosaminoglycans (see above) to three-dimensional nodules displaying osteogenic phenotypic markers, i.e., WV1D1
(9C5) (DSHB) for bone sialoprotein II (Kasugai et al., 1992), MPIII (DSHB) for osteopontine (Gorski et al., 1990), and the von
Kossa procedure (Silber Protein, Chroma-Gesellschaft) for calcium phosphate. In the von Kossa procedure, negative staining
controls were pre-incubated in EGTA, a specific chelator for calcium (Sigma) (Young et al., 1993, 1999, 2001a,b).
32
Intramembranous Bone, was identified as a direct transition from stellate-shaped stem cells to three-dimensional nodules
displaying only osteogenic phenotypic markers, i.e., WV1D1 (9C5) (DSHB) for bone sialoprotein II (Kasugai et al., 1992), MPIII
(DSHB) for osteopontine (Gorski et al., 1990), and the von Kossa procedure (Silber Protein, Chroma-Gesellschaft) for calcium
phosphate. In the von Kossa procedure, negative staining controls were pre-incubated in EGTA, a specific chelator for calcium
(Sigma) (Young et al., 1993, 1999, 2001a,b).
33
Tendon/Ligament, was identified as linear structures with cellular staining for fibroblast specific protein IB10 (Sigma)
(Ronnov-Jessen et al., 1992) and displaying extracellular histochemical staining for type-I collagen (Young et al., 1989c).
34
Dermis, was identified by the presence of interwoven type-I collagen fibers (Young et al., 1989c) interspersed with
spindle-shaped cells staining for fibroblast specific protein IB10 (Sigma) (Ronnov-Jessen et al., 1992) with an extracellular
matrix rich in chondroitin sulfate and dermatan sulfate glycosaminoglycans as assessed by Alcian Blue pH 1.0 staining. In the
latter procedure negative staining controls were digested with chondroitinase-ABC or chondroitinase-AC prior to staining
(Young et al., 1989a,b, 2001a,b).
35
Scar Tissue, was identified as interwoven type-I collagen fibers (Young et al., 1989c) interspersed with spindle-shaped cells
staining for fibroblast specific protein IB10 (Sigma) (Ronnov-Jessen et al., 1992) with an extracellular matrix rich in
chondroitin sulfate glycosaminoglycans as assessed by Alcian Blue pH 1.0 staining. In the latter procedure negative staining
controls were digested with chondroitinase-ABC or chondroitinase-AC prior to staining (Young et al., 1989a,b, 2001a,b).
36
Endothelial Cells, were identified by staining with antibodies P2B1 (DSHB) for CD31-PECAM (Young et al., 2001b), H-Endo
(Chemicon) for CD146 (Gui et al., 1992; Solovey et al., 1997; St. Croix et al., 2000), P8B1 (DSHB) for VCAM (Dittel et al., 1993;
Young et al., 2001b), and P2H3 (DSHB) for CD62e selectin-E (Young et al., 2001b).
37
Hematopoietic Cells, were identified using H-CD34 (Vector) for sialomucin-containing hematopoietic cells (Kishimoto et al.,
1997; Young et al., 2001b), H5A4 (DSHB) for CD11b- granulocytes, monocytes, and Natural Killer cells (August and Hildreth,
2002), H5H5 (DSHB) for CD43 - leukocytes (August and Hildreth, 2002), H4C4 (DSHB) for CD44 - hyaluronate receptor
(August and Hildreth, 2002), Hermes-1 (DSHB) for CD44 - hyaluronate receptor (Butcher, 2002), H5A5 (DSHB) for CD45 - all
leukocytes (August and Hildreth, 2002), and H5C6 (DSHB) for CD63 - macrophages, monocytes, and platelets (August and
Hildreth, 2002).
Endodermal lineage cells were identified as follows.
38
Endodermal Progenitor Cells, were identified with H-AFP (Vector) and R-AFP (NORDIC) for alpha-fetoprotein (Mujoo et al.,
1983).
39
GI Epithelium, was identified with HESA (Sigma) for GI-epithelium (Young, 2004; Young et al., 2004a,b).
40
Liver Oval Cells, were identified with OC2 and OV6 (Hixson) for oval cells, liver progenitor cells, and biliary epithelial cells
(Faris et al., 1991; Gordon et al., 2000).
41
Liver Hepatocytes, were identified with H-1 and H-4 (Hixson) for hepatocyte cell surface marker and hepatocyte cytoplasm,
respectively (Walborg et al., 1985; Faris et al., 1991).
42
Liver Biliary Cells, were identified with OC2, OC3, OC4, OC5, OC10, DPP-IV, and OV6 (Hixson) for biliary epithelial cells,
liver progenitor cells, oval cells, and canalicular cells (Hixson et al., 1984, 1990, 2000; Walborg et al., 1985; Faris et al., 1991;
Gordon et al., 2000).
43
Liver Canalicular Cells, were identified with antibodies H4Ac19 (DSHB), DPP-IV, OV6, and LAP (Hixson) for bile canalic-
ular cells, liver progenitor cells, biliary epithelial cells, and canalicular cell surface protein (Hixson et al., 1984, 1990, 2000;
Hubbard et al., 1985; Walborg et al., 1985; Faris et al., 1991; Gordon et al., 2000).
44
Pancreatic Progenitor Cells, were tentatively identified as three-dimensional structures void of chondrogenic or osteogenic
phenotypic markers. This identity was confirmed by the presence phenotypic markers for pancreatic ductal cells, ␤-Cells,
␣-Cells, and ␦-Cells (Young, 2004; Young et al., 2004a).
45
Pancreatic Ductal Cells, were identified with cytokeratin-19 (Chemicon) to pancreatic ductal cells (Young, 2004; Young et al.,
2004a).
46
Pancreatic ␤-Cells, were identified with YM-PS5088 (Accurate) an antibody to insulin (Young, 2004; Young et al., 2004a).
47
Pancreatic ␣-Cells, were identified with YM-PS087 (Accurate) an antibody to glucagon (Young, 2004; Young et al., 2004a).
48
Pancreatic ␦-Cells, were identified with 11180 (ICN) an antibody to somatostatin (Young, 2004; Young et al., 2004a).

88 YOUNG AND BLACK
We propose that while a majority of cells progress
through the normal embryogenetic developmental se-
quence to form an intact individual, a few cells leave this
genomically preprogrammed pathway to become seques-
tered as reserve precursor cells. For example, a few cells
within the epiblast layer remain as epiblastic-like stem
cells and retain close contacts with the remaining cells as
they differentiate into the three primary germ layer lin-
eages, ectoderm, mesoderm, and endoderm. Similarly, a
few cells from each germ layer remain, respectively, as
ectodermal stem cells, mesodermal stem cells, and
endodermal stem cells, and retain close contacts as the
ectoderm differentiates into surface ectoderm and neural
ectoderm, the mesoderm differentiates into lateral plate
mesoderm, intermediate mesoderm, and paraxial meso-
derm, and the endoderm furthers its differentiation cas-
cade. This process of precursor cell sequestration with
subsequent further differentiation of the majority of the
cells continues to the unipotent progenitor cell and asso-
ciated fully differentiated tissue state. Therefore, every
tissue within the body retains a complement of precursor
cells denoted by its embryological origin. Based on previ-
ous and current studies by ourselves and others (Young,
1977a, 1977b, 1977c, 1983, 2000, 2004; Young et al.,
1983a, 1983b, 1983c, 1983d, 1985, 1989a, 1989b, 1991,
1992a, 1992b, 1993, 1995, 1998a, 1998b, 1999, 2001a,
2001b, 2004a; Lucas et al., 1986, 1988, 1992, 1994a,
1994b, 1995; Lucas and Caplan, 1988; Lucas, 1989; Bow-
erman et al., 1991; Caplan, 1991; Shoptaw et al., 1991;
Gage et al., 1995; Grande et al., 1995; Saito et al., 1995;
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Cornelius et al., 1997; Eglitis and Mezey, 1997; Taylor et
al., 1997; Ferrari et al., 1998; Young and Lucas, 1998;
Gussoni et al., 1999; Petersen et al., 1999; Pittenger et al.,
1999; Alison et al., 2000; Bonner-Weir et al., 2000; Bosch
et al., 2000; Brazelton et al., 2000; Clarke et al., 2000;
Galli et al., 2000; Lagasse et al., 2000; Lee et al., 2000;
Mezey et al., 2000; Ramiya et al., 2000; Sanchez-Ramos et
al., 2000; Woodbury et al., 2000; Deasy et al., 2001;
Jankowski et al., 2001; Reyes and Verfaillie, 2001; Reyes
et al., 2001; Toma et al., 2001; Jiang et al., 2002; McKin-
ney-Freeman et al., 2002; Romero-Ramos et al., 2002) and
Table 1, we would thus propose the model and nomencla-
ture for adult tissue-resident precursor cells shown in
Figure 8. This model and nomenclature are based on the
normal developmental sequence shown in Figure 1.
Previous reports on the identification of adult precursor
cells based their nomenclature on the site of harvest for
the tissue-resident precursor cells, i.e., bone marrow de-
rived, brain derived, skeletal muscle derived, etc. (Eglitis
and Mezey, 1997; Prockop, 1997; Bjornson et al., 1999;
Jackson et al., 1999; Petersen et al., 1999; Brazelton et al.,
2000; Galli et al., 2000; Lagasse et al., 2000; Woodbury et
Fig. 2. Molecular analysis of telomerase activity and Oct-4 expression
in a ␤-Galactosidase-transfected postnatal rat pluripotent epiblastic-like
stem cell clone, designated Rat-A2B2-scl-40 (␤-PPELSC) and a ␤-Galac-
tosidase-transfected postnatal rat pluripotent mesodermal stem cell clone,
designated Rat-A2A2-scl-2PG (␤-PPMSC) (Young et al., 2004a). A: Poly-
acrylamide gel electrophoresis of telomerase activity in pluripotent stem
cells. ␤-PPELSC clone, at 56 passages and 254 population doublings, and
␤-PPMSC clone, at 31 passages and 151 population doublings, were
utilized. Cells were propagated, harvested by trypsin release (Young et al.,
1999) and processed for telomerase activity as described by the manufac-
turer (TRAPeze Assay, Intergen). Lane 1 ⫹, extract of telomerase positive
cells (control), 1 ⫺, extraction buffer, Lane 2 ⫹, test extract of ␤-PPELSC
clone, 2 ⫺, heat inactivated extract of ␤-PPELSC clone; Lane 3 ⫹, test
extract of ␤-PPMSC clone, 3 ⫺, heat inactivated extract of ␤-PPMSC clone.
Note the presence of ladders of bands denoting the presence of telomerase
activity, compare ⫹ lanes 1–3. B: Presence of Oct-4. Oct-4 was detected
by the electrophoretic mobility shift assay using the oligonucleotide 5⬘-
TGTCGAATGCAAATCACTAGA-3⬘ containing the Oct-1 consensus binding
site. A ␤-PPELSC clone at 56 passages and 254 population doublings was
utilized. Cells were thawed, plated at 500 ⫻ 103 cells per 1% gelatinized
T-25 flask in stem cell propagation medium (SCPM), and grown past
confluence. SCPM consisted of 89% (v/v) Opti-MEM, 0.01 mM mercapto-
ethanol (␤ME), 1% antibiotics-antimycotic (ab-am), 10% SS3, at pH 7.4
(Young et al., 2004a). 10% SS3 contained platelet-derived growth factor
(PDGF)-like proliferative and anti-differentiation factor (ADF)-like inhibitory
activities (Young, 2000, 2004; Young et al., 2004a). Cells were harvested by
trypsin release (Young et al., 1999), processed for whole cell extracts as
described (Detn and Latchman, 1993), and aliquoted to 5,000 cell equiva-
lents. Cell aliquots were incubated for 30 min at room temperature with
Lane 1, no competitor; Lane 2, 100-fold excess of unlabelled Oct-1 oligo-
nucleotide; and Lane 3, Oct-4 specific antibody in 20mM Tris, pH 7.5, 4%
glycerol 0.5 mM dithiothreitol, 2␮g poly dIdC. 32P- labeled Oct-1 oligonu-
cleotide (1ng) was added and the mixture incubated for 30 min at room
temperature before electrophoresis through a 5% polyacrylamide gel. After
drying, bands were visualized with a phosphorimager and quantified using
the accompanying software. Two bands that represent binding by mem-
bers of the Oct family of transcription factors were obtained, as shown by
the competition for binding by unlabelled Oct oligonucleotide. C: Densito-
metric analysis of the area contained in the sidebar of the electrophoretic
mobility shift assay in Fig. 2B. Lane 1, solid line; Lane 2, long dashes; and
Lane 3, short dashes. Incubation with Oct-4 specific-antibody substantially
decreased the formation of the upper band, and slightly decreased the
formation of the lower band, indicating the presence of Oct-4.

ADULT STEM CELLS
al., 2000; Deng et al., 2001; Vescovi et al., 2001, Yang et
al., 2002). In contrast, the nomenclature for the precursor
cells shown in Figure 8 is based on the functionality of the
cells, i.e., their proliferation potential with respect to Hay-
flick’s limit and their ability to commit to various tissue
lineages (Fig. 1), rather than their original site of harvest.
This model proposes that two general categories of pre-
cursor cells exist within postnatal animals, including hu-
mans. The proposed general categories are lineage-un-
committed pluripotent stem cells and lineage-committed
progenitor cells. We would also propose that subcategories
exist within each general category of precursor cell and
that each subcategory parallels the function of its respec-
tive counterpart in embryonic development (Fig. 1). The
proposed subcategories for pluripotent stem cells at the
present time are epiblastic-like stem cells, ectodermal
stem cells, surface ectodermal stem cells, neuroectodermal
stem cells, neural tube stem cells, neural crest stem cells,
mesodermal (mesenchymal) stem cells, paraxial mesoder-
mal stem cells, intermediate mesodermal stem cells, lat-
eral plate mesodermal stem cells, and endodermal stem
cells. With time, additional subcategories of pluripotent
stem cells may be identified. The proposed subcategories
for progenitor cells are multipotent cells, tripotent cells,
bipotent cells, and unipotent cells for each respective germ
layer lineage (Fig. 8). Thus, the presence within the tis-
sues of both lineage-uncommitted pluripotent stem cells
and lineage-committed progenitor cells provides for the
continual maintenance and repair of the organism after
birth.
To ascertain the validity or our proposed model and
nomenclature, we have been examining the precursor cell
composition in the connective tissue compartments of
many organs and tissues in nine different species, includ-
ing humans (review, Young, 2004; Young et al., 2004a).
We utilized our in vitro insulin-dexamethasone-ELICA
phenotypic bioassay for the majority of these studies to
assess the identity of the isolated cells. Cellular identity
was based on their proliferation and differentiation poten-
tials. After an initial cell isolate, cells were assayed in
96-well plates in defined testing medium (TM) only, TM
plus 2–5 ␮g/ml insulin, and TM plus 10–10 to 10– 6 M
dexamethasone. The cultures were grown for up to eight
weeks with medium changes three times per week. Cells
were assessed routinely for changes in morphology. After
48 hr to eight weeks the cells were fixed and processed for
qualitative and quantitative ELICAs using our full library
of probes for phenotypic expression markers of embryonic,
ectodermal, mesodermal, and endodermal lineage cells
(see legend to Table 3). Embryonic-like cells and differen-
tiated cells expressed their respective phenotypic markers
after incubation in TM only. Lineage-committed progeni-
tor cells expressed their respective phenotypic differenti-
ation markers after incubation in TM plus insulin. Lin-
eage-uncommitted pluripotent stem cells and lineage-
committed progenitor cells expressed phenotypic markers
of their differentiation potential after incubation in TM
plus dexamethasone. Therefore, to obtain a true measure
of pluripotent stem cell content, it was necessary to sub-
tract the expressed differentiation markers in TM plus
insulin from those generated by TM plus dexamethasone.
These studies demonstrated that approximately 50% of
the precursor cells residing in a tissue-specific connective
tissue compartment were lineage-committed progenitor
cells specific for that particular tissue. Approximately 40%
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
89
of the precursor cells were lineage-committed unipotent,
bipotent, tripotent, and multipotent progenitor cells for
other ectodermal, mesodermal, and endodermal tissue lin-
eages. Approximately 9% of the precursor cells were plu-
ripotent stem cells belonging to the ectodermal, mesoder-
mal, or endodermal cell lineages. And approximately 1% of
the precursor cells were pluripotent epiblastic-like stem
cells, capable of forming any somatic cell of the body.
Using the precursor cells derived from skeletal muscle as
an example, approximately 50% of the reserve precursor
cells located in skeletal muscle consist of progenitor cells
committed to the myogenic lineage. Approximately 40%
are progenitor cells committed to other lineages (i.e., adi-
pogenic, fibrogenic, osteogenic, chondrogenic, endothelio-
genic, hematopoietic, neurogenic, gliogenic, hepatogenic,
etc.). Approximately 9% are pluripotent germ layer stem
cells (i.e., pluripotent ectodermal stem cells, PPMSCs, and
pluripotent endodermal stem cells). And approximately
1% are pluripotent epiblastic-like stem cells. Precursor
cells from the connective tissue compartments of such
diverse tissues as dermis of the skin, bone marrow, brain,
adipose tissue (fat), perichondrium, periosteum, and vis-
cera appear to follow the same pattern of progenitor cell
and pluripotent stem cell distribution as precursor cells
derived from the connective tissue compartments of skel-
etal muscle (Young et al., 1993, 1995, 1998a, 2001a,
2001b, 2004a; Young, 2000, 2004). Others have noted the
presence of precursor cells within the tissues of animals,
including humans, with similar capabilities as those de-
scribed above (Table 1).
DIFFERENTATION OF PLURIPOTENT STEM
CELLS VS. TRANSDIFFERENTIATION OF
PROGENITOR CELLS
Researchers have recently reported that precursor cells
derived from one organ are being reprogrammed to form
tissues of another organ. For example, precursor cells
derived from skeletal muscle were initially reported to
form blood (Jackson et al., 1999). Precursor cells derived
from bone marrow have been reported to form neurons
and neural supportive tissues (Eglitis and Mezey, 1997;
Brazelton et al., 2000; Woodbury et al., 2000), hepatic oval
cells (Petersen et al., 1999; Lagasse et al., 2000; Theise et
al., 2000), and muscle cells (Ferrari et al., 1998; Gussoni et
al., 1999). Precursor cells derived from neuronal tissues
have been reported to form blood elements (Bjornson et
al., 1999; Vescovi et al., 2001) and muscle cells (Clarke et
al., 2000; Galli et al., 2000; Tsai and McKay, 2000). And
precursor cells derived from the liver have been reported
to form pancreatic islet cells (Yang et al., 2002). These
investigators proposed the theory of transdifferentiation,
which involves the genetic reprogramming of progenitor
cells from one cell lineage to form differentiated cells be-
longing to another lineage. However, none of these studies
addressed either the identity or differentiative capability
of their precursor cells prior to experimentation. There-
fore, the theory of transdifferentiation amounts to more of
an assumption than a proven theory.
We believe that it is necessary to understand the histo-
logical makeup of any given tissue and its inherent em-
bryological origin (Fig. 1) to realize the potential popula-
tions of precursor cells that may reside in that tissue. For
example, the organ of skeletal muscle is composed of skel-
etal muscle fibers of myotomal somitic or lateral plate
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
YOUNG AND BLACK

Figure 3.
90

ADULT STEM CELLS
somatic mesodermal origin, depending on the location of
the muscle within the body. The endomysial connective
tissue surrounding individual myofibers, the perimysial
connective tissue surrounding muscle fascicles, and the
epimysial connective tissues surrounding the entire mus-
cle are of sclerotomal somitic or lateral plate somatic me-
sodermal origin, depending on the location of the muscle.
The motor end plates and innervations to the muscle are
of neural crest origin. And the vasculature, blood vessels
and lymphatic vessels, arise from mesoderm and are thus
of somitic or lateral plate somatic mesodermal origin, de-
pending on location of the muscle. The blood vessels also
contain circulating differentiated blood cells and blood
stem cells derived from lateral plate splanchnic meso-
derm. Based on the histology and embryological origin of
the tissue, we would expect to find the following precursor
cells within the organ of skeletal muscle: pluripotent epi-
blastic-like stem cells (representing all tissues within the
organ of skeletal muscle), PPMSCs (skeletal muscle, con-
nective tissue, and vasculature), pluripotent ectodermal
stem cells (motor end plates, peripheral nerves and their
supportive glial cells), pluripotent neural crest cells (mo-
tor end plates, peripheral nerves and their supportive
glial cells), and multipotent, tripotent, bipotent, and
unipotent progenitor cells for each of the above-mentioned
tissue types, i.e., skeletal muscle myofibers, connective
tissue coverings, vessels, blood, blood stem cells, and ner-
vous tissues.
We (Young, 1977a, 1977b, 1977c, 1983, 2000, 2004;
Young et al., 1983a, 1983b, 1983c, 1983d, 1985, 1989a,
1989b, 1991, 1992a, 1992b, 1993, 1995, 1998a, 1998b,
1999, 2001a, 2001b, 2004a, 2004b; Lucas et al., 1993,
1995, 1996a, 1996b; Pate et al., 1993; Rogers et al., 1995;
Fig. 3. Rat clone (A2A2) of postnatal pluripotent mesodermal stem
cells (PPMSC) (A, B, D, G, I, N) and human pluripotent mesodermal stem
cell lines (CD10⫹, CD13⫹, CD34⫹⫹, CD56⫹, CD90⫹, MHC-I⫹), i.e., hu-
man NHDF2 cells, derived from a dermal biopsy specimen taken from a
36 year-old human female, at 80 cell doublings (C, E, F, H, K-M, P-T) and
PAL3 cells, derived from 67 year old human male skeletal muscle biopsy
specimen, at 150 cell doublings (J, O) demonstrating mesodermal mor-
phologies. Cells were grown in stem cell medium (SCM), testing medi-
um-6 (TM-6), or TM-6 with 10⫺6 M Dexamethasone (Dex) for 1 to 56
days. SCM consisted of 89% (v/v) Eagle’s MEM with Earle’s salts, 1%
antibiotics (ab), 10% SS3, pH 7.4. Selected serum-3 (SS3) contained
PDGF-like proliferative and ADF-like inhibitory activities (Young, 2000).
TM-6 consisted of 89, 94, or 98% (v/v) Opti-MEM with 0.01 mM ME; 10,
5, or 1% SS9; 1% ab-am, 2 ␮g/ml insulin, at pH 7.4. SS9 contained
inductive activities resembling those of skeletal muscle morphogenetic
protein (Sk-MMP), adipocyte morphogenetic protein (AMP), and bone
morphogenetic protein-2 (BMP-2) (Young, 2000). Cultures were grown in
SCM (A), TM-6 only (B), TM-6 with 10⫺6 M dexamethasone (Dex) (D, G,
I, N), TM-6 with 1%-SS9 (C, E, F, H, K-M, P-T), TM-6 with 10%-SS9 (J),
TM-6 with 1%-SS3 (O) for 1 day (A), 14 days (E), 28 days (M), 42 days (D,
G, J, I, N, O), and 56 days (C, F, H, K, L, P-T). Morphologies and
histochemical and immunochemical staining as noted (see also legend
to Table 3). Photographed with phase contrast (A-C) and brightfield (D-T)
microscopy, original magnifications ⫻ 200 (A, B), ⫻ 100 (C-L, N, P-R, T),
⫻ 50 (O), ⫻ 40 (S), ⫻ 25 (M). (From Young et al., 2001a, Clonogenic
analysis reveals reserve stem cells in postnatal mammals. I. Pluripotent
mesenchymal stem cells. Anatomical Record 263:350-360. Reprinted
with permission from Wiley-Liss, Inc. (A, B, D, G, I, N).; Young et al.,
2001b, Human reserve pluripotent mesenchymal stem cells are present
in the connective tissues of skeletal muscle and dermis derived from
fetal, adult, and geriatric donors. Anat. Rec. 264:51-62, Reprinted with
permission from Wiley-Liss, Inc. (C, E, F, H, J-M, O-T)). A: Small mono-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
91
Dixson et al., 1996; Warejcka et al., 1996; Young and
Lucas, 1998; Romero-Ramos et al., 2002) and others
(Reyes and Verfaillie, 2001; Jiang et al., 2002; Zhao et al.,
2003) (see also Table 1) have observed that many tissues
and organs contain a wide variety of precursor cells. The
precursor cells observed include pluripotent epiblastic-
like stem cells, pluripotent ectodermal stem cells, PPM-
SCs, and pluripotent endodermal stem cells; and multipo-
tent, tripotent, bipotent, and unipotent progenitor cells.
These precursor cells can be isolated from normal healthy
tissues using numerous techniques, i.e., mechanical dis-
ruption of the tissues, outgrowth, enzymatic release, dif-
ferential plating, etc. Then as a cell isolate the precursor
cells can be further segregated from differentiated cells by
cryopreservation, cell sorting, differential centrifugation
with and without percoll/ficoll gradients, log-phase expan-
sion, etc.
In an ongoing series of studies (Young et al., 1993,
1998b; Young, 2000, 2004; unpublished observations) we
have been examining the potential for transdifferentiation
in lineage-committed progenitor cell populations. We pos-
tulated that if the theory of transdifferentiation were a
reality, then we could induce a unipotent lineage-commit-
ted progenitor cell clone to exhibit phenotypic expression
markers for an alternate tissue lineage. In our first series
of experiments we utilized five separate unipotent progen-
itor cell clones for the fibrogenic, myogenic, adipogenic,
chondrogenic, and osteogenic lineages, respectively. Next,
we incubated these clones individually for seven days with
recombinant or novel inductive factors outside their re-
spective tissue lineages, removed the inductive factor, and
then incubated the cells with insulin to accelerate pheno-
typic expression. The inductive factors utilized were fibro-
nucleated cells with large ratios of nucleus to cytoplasm. B: Single layer
of nondescript contact-inhibited cells. C: Two linear structures (arrows)
containing multiple nuclei. D: Dark structures stained with antibody to
sarcomeric myosin (MF-20) (arrow). Majority of unstained cells in back-
ground are adipocytes (fat cells) (asterisks). E: Mononucleated cells
staining intracellularly for smooth muscle alpha-actin (IA4). F: Binucle-
ated cell co-staining intracellularly for sarcomeric myosin (MF-20) and
smooth muscle alpha-actin (IA4). G: Cells with multiple Oil Red-O
stained intracellular vesicles indicative of saturated neutral lipid-contain-
ing adipocytes (arrow). H: Mononucleated cell staining intracellularly for
type-II collagen (CIIC1). I: Aggregating nodule of cells (single arrow) with
pericellular matrix halos staining with antibody to type-IX collagen (D1-
9). Also note aggregation of unstained adipocytes (double arrows) and
individual cells (asterisk) stained with antibody to smooth muscle alpha-
actin (1A4). J: Nodule stained for chondroitin sulfate and keratan sulfate
glycosaminoglycans (Alcian Blue, pH 1.0). K: Mononucleated cells stain-
ing intracellularly for osteopontine (MP111). L: Mononucleated cells
staining intracellularly for bone sialoprotein II (WV1D1). M: Aggregation
of cells with pericellular matrix halos. N: Aggregating nodule of cells
overlaid with horse shoe-shaped extracellular matrix staining with anti-
body to bone sialoprotein (WV1D1) (white asterisk). Unstained refractile
intercellular vesicles belong to adipocytes (single arrows). A diagonally
oriented smooth muscle cell stained with an antibody to smooth muscle
alpha-actin (1A4) (double arrows) is located in the upper left-hand corner
of photograph. O: Nodules stained for calcium phosphate (von Kossa).
P: Mononucleated cells stained for fibroblast specific protein (1B10).
Q: Mononucleated cells stained for human endothelial cell surface
marker (P1H12). R: Mononucleated cells stained for peripheral cell ad-
hesion molecule (P2B1). S: Mononucleated cells stained for vascular cell
adhesion molecule (P8B1). T: Mononucleated cells stained for E-selectin
(P2H3).

92 YOUNG AND BLACK
blast morphogenetic protein (FMP) to induce fibrogenesis,
adipocyte morphogenetic protein (AMP) to induce adipo-
genesis, BMP-2 to induce chondrogenesis and osteogene-
sis, and Sk-MMP to induce skeletal myogenesis. In all
cases examined, the tissue-specific progenitor clones
maintained their original lineage commitment program-
ming and exhibited tissue-specific phenotypic expression
markers, even in the presence of inductive factors outside
their respective lineages, i.e., FMP, AMP, BMP-2, or Sk-
MMP. These results suggested that once a cell is commit-
ted to a particular tissue lineage, it would not change its
genetic programming, i.e., revert to a more primitive state
and express markers for a different tissue lineage.
In a second series of experiments we induced the forma-
tion of unipotent lineage-committed progenitor cells from
a lineage-uncommitted PPMSC clone using the same tis-
sue-specific lineage induction factors as above (i.e., FMP,
AMP, BMP-2, Sk-MMP). After establishment of the unipo-
tent progenitor cell lines, we repeated the above experi-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
ment utilizing these same specific inductive factors with a
seven-day incubation schedule, removal of the inductive
factor, and then incubation of the cells with insulin to
accelerate phenotypic expression. In all experiments the
induced unipotent progenitor cell lines maintained their
originally induced lineage commitment programming.
Again, these results suggested that once a cell is commit-
ted to a particular tissue lineage, it would not change its
genetic programming.
In a third series of experiments we mixed a clone of
lineage-uncommitted pluripotent stem cells with one or
more clones of lineage-committed progenitor cells. We
then incubated these mixed cultures with one or more
tissue-specific induction factors (same as above) with and
without insulin. The results demonstrated that the pro-
genitor cells were responsive to insulin and would express
their lineage-committed phenotype in the presence of in-
sulin or their respective lineage induction factor. In con-
trast, the lineage-uncommitted pluripotent stem cell clone
responded to the lineage-specific induction factor, but not
to insulin. For example, a myogenic clone was mixed with
a chondrogenic clone and a pluripotent stem cell clone.
The mixed culture was then incubated with FMP and
insulin, FMP alone, and insulin alone. The mixed culture
incubated with FMP and insulin demonstrated phenotypic
expression markers for myogenesis, chondrogenesis, and
fibrogenesis. The mixed culture incubated with FMP alone
demonstrated only phenotypic expression markers for fi-
brogenesis. The mixed culture incubated with insulin
alone demonstrated phenotypic expression markers for
myogenesis and chondrogenesis. These results suggested
that the myogenic and chondrogenic clones within the
mixed culture were responding to insulin, while the plu-
ripotent stem cell clone in the mixed culture was respond-
ing to FMP.
Fig. 4. Human (CD10⫹, CD66e⫹) (A, C, E- G, I-K) and Rat clone
(A2B2-scl-40) (B, D, H, L) postnatal pluripotent epiblastic-like stem cells
(PPELSC) demonstrating embryonic markers. PPELSC lines were grown
for 24 hours to 8 weeks in serum-free testing medium (TM) only, TM with
10% SS3, or TM with 2 ␮g/ml insulin. TM consisted of 99% (v/v)
Opti-MEM, 0.01 mM ␤ME, 1% ab-am, pH 7.4. 10% SS3 contains
proliferative activity resembling that of PDGF and inhibitory activity
resembling that of ADF (Young, 2000, 2004; Young et al., 2004a). In this
particular series a human (CD10⫹, CD66e⫹) PPELSC line and a rat clone
(A2B2-scl-40) PPELSC line were incubated in TM only (A, B), TM with
10% SS3 (C, D), or TM with insulin (E-L) for 24 hours (A, B) or seven days
(C-L). Morphologies and immunochemical staining as noted (see also
legend to Table 3). Photographed with phase contrast (A-D) and bright-
field microscopy (E-L). Original magnifications, ⫻ 400 (A, E, F), ⫻ 200 (B,
D, G-K) and ⫻ 100 (C, L). A: Very small cells with high ratios of nucleus
to cytoplasm. Stellate shape denotes an attached cell, linear shape is
indicative of mobile cell. B: Very small cells with high ratios of nucleus to
cytoplasm. Linear shape is indicative of mobile cell. C: Multilayered
confluent cells maintaining stellate morphology. D: Multilayered conflu-
ent cells maintaining stellate morphology. E: Moderate to heavy staining
for stage specific embryonic antigen-1 (SSEA-1). F: Moderate staining
for stage specific embryonic antigen-3 (SSEA-3). G: Moderate to heavy
staining for stage specific embryonic antigen-4 (SSEA-4). H: Moderate
to heavy staining for stage specific embryonic antigen-4 (SSEA-4).
I: Moderate to heavy staining for human carcinoembryonic antigen
(HCEA). J: Moderate to heavy staining for human carcinoembryonic
antigen (CD66e). K: Moderate staining for carcinoembryonic antigen cell
adhesion molecule-1 (CEA-CAM1). L: Moderate to heavy staining for
carcinoembryonic antigen cell adhesion molecule-1 (CEA-CAM1).

ADULT STEM CELLS
Fig. 5. A human (CD10⫹, CD66e⫹) PPELSC line (A, C, E, G, H, J, L,
M, O, S) and rat clone (A2B2-scl-40) PPELSC line (B, D, F, I, K, N, P-R)
were induced to express ectodermal (neuroectodermal and surface ec-
todermal) phenotypic expression markers. The PPELSC lines were
plated at 103 cells per well in 1% gelatin-coated 96-well plates and
grown for 24 hours to 8 weeks in testing medium (TM) only; TM with 10⫺6
to 10⫺10 M Dex; TM with 10, 5, or 1% SS12; or TM with 10⫺6 to 10⫺10
M Dex and 10, 5, 3, or 1%-SS12. TM consisted of 89, 94, 96, or 98%
(v/v) Opti-MEM with 0.01 mM ␤ME, 1% ab-am, 2 ␮g/ml insulin, at pH
7.4. Five, 3, and 1% SS12 contains ectodermal inductive activities. In
this particular series human cells were grown for 7 days in TM with 10⫺6
M Dex and 1% SS12. Morphologies and immunochemical staining as
noted (see also legend to Table 3). Photographed with brightfield mi-
croscopy, original magnifications, ⫻ 200 (A, C-H, J, L-O, S), ⫻ 100 (B, I,
P-R), ⫻ 40 (K). A: Mononucleated cells moderately to heavily stained for
neural precursor cell expression marker (FORSE-1). B: Mononucleated
cells heavily stained for neural precursor cell expression marker (FORSE-
1). C: Mononucleated cells showing heavy intracellular staining for neu-
rofilaments (RT-97). D: Mononucleated cells showing moderate to heavy
intracellular staining for neurofilaments (RT-97). E: Mononucleated cells
showing moderate to heavy intracellular staining for neurons (8A2).
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
93
F: Mononucleated cells showing moderate to heavy intracellular staining
for neurons (8A2). G: Mononucleated cells showing moderate to heavy
intracellular staining for nestin (HNES). H: Mononucleated cell showing
heavy intracellular staining for nestin (MAB353). I: Mononucleated cells
showing heavy intracellular staining for neuronal nestin (Rat-401).
J: Mononucleated cells showing low to moderate intracellular staining
for b-tubulin-III (T8660). K: Mononucleated cells showing moderate to
heavy intracellular staining for b-tubulin-III (T8660). L: Mononucleated
cells showing moderate to heavy intracellular staining for neuroglia (oli-
godendrocytes and astroglia) (CNPase). M: Mononucleated cell showing
heavy intracellular staining for oligodendrocytes (Rip). N: Mononucleated
cell showing heavy intracellular staining for oligodendrocytes (Rip).
O: Mononucleated cells showing heavy intracellular staining for neuronal
expression marker (S-100). P: Mononucleated cells showing moderate
intracellular staining for neuronal expression marker (S-100). Q: Mono-
nucleated cells showing moderate to heavy intracellular staining for
neuronal vimentin for radial cells and radial glial cells (40E-C). R: Mono-
nucleated cells showing moderate to heavy intracellular staining for
ganglion cells (TuAg1). S: Mononucleated cells showing low to moderate
staining for keratinocytes (VM-1).

94 YOUNG AND BLACK
Fig. 6. Human postnatal pluripotent epiblastic-like stem cell line
(CD10⫹, CD66e⫹) (B, C, D, H, M, N, O, P) and postnatal rat pluripotent
epiblastic-like stem cell clone (A2B2-scl-40) (A, E, F, G, I, J, K, L) induced
to express mesodermal phenotypic expression markers. Multiple
PPELSC lines were plated at 103 cells per well in 1% gelatin-coated
96-well plates and grown for 24 hours to 8 weeks in testing medium (TM)
only; TM with 10⫺6 to 10⫺10 M Dex; TM with 10, 5, 3, or 1% SS9 or 1%
SS3; or TM with 10⫺6 to 10⫺10 M Dex and 10, 5, 3, or 1% SS9 or 1%
SS3. TM consisted of 89, 94, 96, or 98% (v/v) Opti-MEM with 0.01 mM
␤ME with 1% ab-am, 2 ␮g/ml insulin, at pH 7.4. Ten, 5, 3, and 1% SS9
contains inductive activities resembling that of Sk-MMP, AMP, BMP-2,
and endothelial inductive activity, while 1% SS3 contains inductive
activity resembling that of Sm-MMP (Young, 2004; Young et al., 2004a).
In this particular series PPELSCs were grown for 1 week (A, B, C, D, L-P),
2 weeks (E, H), or 4 weeks (F, G, I- K) in TM with 10⫺8 M Dex plus insulin
plus 1% SS9 (A, B, L-P), 10⫺8 M Dex plus insulin plus 1% SS3 (C), or
10⫺7 M Dex plus insulin plus 3% SS9 (E- K). Morphologies and immu-
nochemical staining as noted (see also legend to Table 3). Photographed
with brightfield microscopy; original magnifications, ⫻ 200 (C, L, N), ⫻
160 (O), ⫻ 100 (A, B, D, E, F, G, H, I, K, M, P), ⫻ 40 (J). A: Mononucleated
cells showing heavy intracellular staining for myogenin (F5D). B: Mono-
nucleated and binucleated cells and multinucleated linear and branched
structures showing moderate to heavy intracellular staining for anti-
skeletal muscle fast myosin (MY-32). C: Mononucleated cells showing
Throughout all our studies trying to prove the validity of
transdifferentiation we were never able to repeat trans-
differentiation results utilizing known populations of pre-
cursor cells with known differentiation potentials. There-
fore, based on the above experiments, we would propose
that differentiated progenitor cells do not undergo trans-
differentiation or dedifferentiation. We now believe that
the experimental results that appear to require transdif-
ferentiation or dedifferentiation of progenitor cells to oc-
cur during adult tissue restoration when progenitor cells
derived from one organ are reprogrammed to form tissues
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
heavy intracellular staining for smooth muscle ␣-actin (IA4). D: Mono-
nucleated and binucleated cells showing moderate intracellular staining
for cardiotin (cardiac myocytes, MAB 3252). E: Mononucleated cells
demonstrating heavy intracellular staining for bone sialoprotein II
(WV1D1). F: Nodule of cells demonstrating extracellular staining for
osteopontine (MP111). G: Nodule of cells demonstrating extracellular
staining for calcium phosphate using the von Kossa procedure (vK).
H: Mononucleated cells with heavy intracellular staining for cartilage-
specific collagen pro type-II (CIIC1). I: Single nodule of cells demonstrat-
ing moderate to heavy extracellular staining for cartilage-specific colla-
gen type-II (HC-II). J: Three nodules demonstrating heavy extracellular
staining for cartilage-specific collagen type-IX (D1-9). K: Two nodules
demonstrating extracellular staining for sulfated glycosaminoglycan
chains of proteoglycans (Alcian Blue-O, pH 1.0). L: Mononucleated cells
with moderate to heavily stained intracellular vesicles demonstrating
saturated neutral lipids (Oil Red-O), indicative of adipocytes. M: Three-
dimensional tubular structure demonstrating moderate to heavy staining
for CD146, endothelial cells (H-endo). N: Mononucleated cell demon-
strating heavy staining for VCAM, vascular (endothelial) cell adhesion
molecule (P8B1). O: Mononucleated cells demonstrating low to moder-
ate staining for CD62e, (endothelial) selectin-E (P2H3). P: Mononucle-
ated cells demonstrating moderate staining for CD44, hyaluronate re-
ceptor (H4C4).
of another organ (review, Forbes et al., 2002; Poulsom et
al., 2002; Tsai et al., 2002) are actually due to contamina-
tion of the tissue isolate by unrecognized progenitor cells
and/or pluripotent stem cells that are stimulated to form
new progenitor cells of a different tissue lineage. For ex-
ample, we propose that pluripotent stem cells (and/or he-
matopoietic progenitor cells) present in skeletal muscle
could give rise to the blood cells initially reported by
Jackson et al. (1999). Indeed, a subsequent analysis by the
same research group (McKinney-Freeman et al., 2002), in
an elegant study demonstrated that CD45⫹ (hematopoi-

ADULT STEM CELLS
Fig. 7. Human (CD10⫹, CD66e⫹) postnatal pluripotent epiblastic-like
stem cell line (B, D, G, N, O) and rat postnatal pluripotent epiblastic-like
stem cell clone, A2B2-scl-40 (A, C, E, F, H-M, P) (PPELSCS) induced to
express endodermal phenotypic markers. Multiple PPELSC lines were
plated at 103 cells per well in 1% gelatin-coated 96-well plates and
grown for 24 hours to 8 weeks in testing medium (TM) only; TM with 10⫺6
to 10⫺10 M Dex; TM with 15 or 10% SS12; or TM with 10⫺6 to 10⫺10 M
Dex and 15 or 10% SS12. Testing medium consisted of 74 or 89% (v/v)
Opti-MEM with 0.01 mM ␤ME with 1% ab-am, 2 ␮g/ml insulin, at pH 7.4.
15 and 10% SS12 contains endodermal inductive activities. In this
particular series PPELSCs were incubated for 1 week (A, D, E, G, N, O),
2 weeks (F, K, L, Q) 3 weeks (I, J, M), 4 weeks (B, P), or 5 weeks (C, H)
in TM with 15% SS12 and 10⫺6 M Dex. Morphologies and immuno-
chemical staining as noted (see also legend to Table 3). Photographed
with brightfield microscopy, original magnifications, ⫻ 200 (A), ⫻ 100
(C-P). A: Mononucleated and binucleated cells showing intense intra-
cellular staining for rat-specific alpha-fetoprotein (RAFP). B: Mononucle-
ated cell showing heavy intracellular staining for gastro-intestinal epi-
thelium (HESA). C: Nodular aggregation showing moderate intracellular
staining for pro-insulin in ␤-cells of endocrine pancreas (YM-PS5088).
D: Mononucleated cells showing moderate to intense intracellular stain-
ing for glucagon in ␣-cells of endocrine pancreas (YM-PS087). E: Indi-
vidual diffuse mononucleated cells showing moderate to intense intra-
etic) stem cells residing in skeletal muscle gave rise to
blood cells, whereas the CD45– (nonhematopoietic) stem
cells residing in skeletal muscle formed muscle. However,
these investigators did not examine the CD45– stem cells
of skeletal muscle to determine their full differentiation
potential, i.e., whether they would form other cell/tissue
types in addition to skeletal muscle. Several possibilities
exist. The CD45– stem cells could be progenitor cells com-
mitted to the myogenic lineage. Alternatively, the CD45–
stem cells could be PPMSCs or pluripotent epiblastic-like
stem cells. The CD45– cells could also be a combination of
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
95
cellular staining for somatostatin in ␦-cells of endocrine pancreas
(11180). F: Cellular aggregation showing moderate to intense intracellu-
lar staining for ductal cells of exocrine pancreas (CK-19). G: Mononucle-
ated cells showing light to moderate intracellular staining for bile cana-
licular cells of liver (HA4c19). H: Nodule showing heavy intracellular
staining for progenitor cells, biliary epithelial cells, and oval cells of liver
(OC2). I: Cellular aggregation and individual diffuse mononucleated cells
showing moderate to heavy intracellular staining for progenitor cells and
biliary epithelial cells of liver (OC3). J: Cellular aggregation and individual
diffuse mononucleated cells showing moderate to intense intracellular
staining for progenitor cells and biliary epithelial cells of liver (OC4).
K: Diffuse mononucleated cells showing moderate to heavy intracellular
staining for progenitor cells and biliary epithelial cells of liver (OC5).
L: Diffuse mononucleated cells showing moderate to intense intracellular
staining for progenitor cells and biliary epithelial cells of liver (OC10).
M: Diffuse and aggregated cells showing moderate to intense intracel-
lular staining for cytoplasm of liver hepatocytes (H.4). N: Mononucleated
cells showing moderate to intense staining for liver hepatocyte cell
surface marker (H.1). O: Mononucleated cells showing light to moderate
to intense staining for progenitor cells, canalicular cells, and biliary
epithelial cells of liver (DPP-IV). P: Diffuse cells showing moderate to
heavy intracellular staining for biliary epithelial cells, oval cells, hepato-
cyte canalicular cells of liver (OV6).
all three types of precursor cell, since all of these types of
cells are CD45– by flow cytometric analysis (Young, 2000,
2004; Young et al., 2001b, 2004a).
Along a similar vein as above, Petersen et al. (1999) and
Yang et al. (2002) described an adult progenitor cell from
bone marrow that would form liver hepatocytes and an
adult hepatic oval (stem) cell from liver that would form
pancreatic islets (pancreatic endocrine hormone-produc-
ing cells). In both instances they failed to characterize
their identified stem cells for pluripotency prior to their
experimental studies. They then used the theory of trans-

96 YOUNG AND BLACK
Fig. 8. Flow chart of adult stem cell hierarchy and nomenclature,
from most primitive (top) to most differentiated (bottom). Adult stem cells
are divided into two major categories: pluripotent stem cells (PPSCs)
and progenitor cells (ProgCs). Characteristics of each category are as
follows. PPSCs are lineage-uncommitted, telomerase positive, and have
extensive capabilities for self-renewal. They are composed of at least the
following categories, i.e., pluripotent epiblastic-like stem cells
(PPELSCs), pluripotent germ layer lineage stem cells and subcategories
of pluripotent germ layer lineage stem cells, such as pluripotent ecto-
dermal stem cells (PPEctoSCs), pluripotent surface ectodermal stem
cells (PPSurfEctoSCs), pluripotent neuroectodermal stem cells (PPNeu-
roEctoSCs), pluripotent neural tube stem cells (PPNT-SCs), pluripotent
neural crest stem cells (PPNC-SCs), pluripotent mesodermal stem cells
differentiation to explain the formation of tissues other
than that from which the cells were isolated. Based on our
studies (above, Young et al., 1998a, 1998b, 2001b, 2004a;
Young, 2000, 2004) and those of others (Reyes and Ver-
faillie, 2001; Jiang et al., 2002), we believe that the start-
ing populations of tissue-specific precursor cells in the
studies by Goodell’s group (Jackson et al., 2000; McKin-
ney-Freeman et al., 2002) and Petersen’s group (Petersen
et al., 1999; Yang et al., 2002) could also be explained by
the presence of pluripotent stem cells (and/or lineage-
committed progenitor cells for other tissue lineages) as
resident precursor cell populations within the respective
differentiated tissues examined. Thus, it is not necessary
to invoke the concept of transdifferentiation to explain the
results obtained by these investigative groups.
We suggest that future experiments addressing the is-
sue of transdifferentiation of adult precursor cells should
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
(PPMSCs), pluripotent paraxial mesodermal stem cells (PPPMSCs), plu-
ripotent intermediate mesodermal stem cells (PPIMSCs), pluripotent
lateral plate mesodermal stem cells (PPLPMSCs), and pluripotent
endodermal stem cells (PPEndoSCs). PPELSCs have the capacity to
form any somatic cell of the body and display embryonic phenotypic
markers in the lineage-uncommitted state. Pluripotent germ layer stem
cells have the capacity to form any cell type within their respective germ
layer lineage and display lineage-specific phenotypic markers. ProgCs
are lineage-committed, e.g., committed to forming cells exclusive to
particular germ layer lineages, and conform to Hayflick’s Limit of 50-70
population doublings before cell senescence and cellular death. They
are composed of multipotential, tripotential, bipotential, and unipotential
precursor cells for their respective germ layer lineages.
begin with the characterization of the composition and
complete differentiation potentials of the precursor cells
utilized. For those investigators that still maintain that
transdifferentiation is a valid hypothesis, we would sug-
gest a thorough characterization of the differentiation po-
tential of their cells prior to the onset of experimentation.
Unless this information is made available, it is very diffi-
cult to interpret the results of such experiments. Due to
the ubiquitous locations of identified precursor cells, we
would also suggest that future experiments addressing
the issue of precursor cell function begin with a complete
characterization of the isolated precursor cells, including
their inherent proliferation potential with respect to Hay-
flick’s limit and their inherent differentiation potential
with respect to the embryogenetic developmental se-
quence. The designated nomenclature for the isolated pre-
cursor cells should then be based on the functionality of

ADULT STEM CELLS
the cells, i.e., proliferation and differentiation potentials,
rather than their location within the tissue.
TISSUE ENGINEERING
With respect to the treatment of trauma and disease,
adult (postnatal) precursor cells can be very useful for
therapies involving autologous transplantation. Precursor
cells can be isolated from newborn to geriatric individuals,
including patients awaiting treatment. Use of autologous
precursor cells circumvents the inherent morbidity and
mortality associated with HLA mismatches that require
the use of immunosuppressant drugs. Adult pluripotent
stem cells can be harvested from a small biopsy of easily
accessible tissues, such as skeletal muscle or dermis. Plu-
ripotent stem cells are telomerase positive, indicating that
vast quantities of cells can be produced from a few har-
vested cells. Pluripotent stem cells can be stored for long
periods of time with minimal loss of cell viability, pluripo-
tentiality, and function. Progenitor cells derived from plu-
ripotent stem cells mimic native progenitor cells with re-
spect to loss of differentiative potential with increased
lineage commitment and adherence to Hayflick’s limit of
50 –70 population doublings before programmed cell se-
nescence and cell death. Pluripotent ectodermal stem cells
have the potential to form neurons, neuroglia, peripheral
nerves, neuroendocrine cells, epidermis, and other cells of
the ectodermal germ layer lineage. PPMSCs have the
potential to form skeletal muscle, smooth muscle, cardiac
muscle, fat, cartilage, bone, tendons, ligaments, dermis,
connective tissues, blood vessels, hematopoietic cells, and
other cells of the mesodermal germ layer lineage. And
pluripotent endodermal stem cells have the potential to
form hepatocytes, biliary cells, canalicular cells, pancre-
atic islet cells, exocrine pancreatic cells, gastrointestinal
epithelial cells, nonpancreatic endocrine cells, and other
cells of the endodermal germ layer lineage. Further, adult
pluripotent epiblastic-like stem cells have the potential to
form cells from all three primary germ layer lineages, i.e.,
ectoderm, mesoderm, and endoderm. Additional studies
are planned or currently under way (Young et al., 2004a)
to determine whether adult reserve pluripotent stem cells
can be used in therapeutic treatments for various condi-
tions such as Parkinson’s disease, stroke, spinal cord re-
pair, blood transfusions, bone marrow transplants, diabe-
tes mellitus and other endocrine diseases, renal failure,
arthritis, muscular dystrophy, vascular ischemia, and
myocardial infarctions.
ACKNOWLEDGMENTS
I especially thank Paul A. Lucas for the generous ex-
change of ideas, reagents, and unpublished observations. I
thank my collaborators, co-authors, and their technical
assistants for their insight and work ethic. We thank Julie
A. Floyd and Nicholas L. Henson for cell culture assis-
tance; Kris Detmer and Angela Thompson for assistance
with the telomerase and Oct-4 analyses; Cecile Duplaa
and Catherine Moreau for ␤-galactosidase transfections;
and Nicholas L. Henson and John Knight for photographic
assistance. The MyoD1 clone, maintained in the expres-
sion vector pVZCII-␣3 scribe, and the myogenin cDNA
probe, subcloned into Bluescribe pEMSV-␣2 M13, were
generous gifts from Dr. W.E. Wright. Bovine nasal septum
A1D1 proteoglycan and the 5D4 anti-keratan sulfate an-
tibody were the kind gifts of Dr. J.H. Kimura. The rh-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
97
BMP-2 and rhBMP-4 were the generous gifts of I.K. Mout-
satsos, Genetics Institute, Inc., Cambridge, MA. The
PDECGF, IGF-II, IGF-I, PDGF-AA, and PDGF-BB were
generous gifts from G.F. Pierce, Amgen, Thousand Oaks,
CA. Sk-MMP, Sm-MMP, AMP, FMP, SIF, and ADF were
the generous gifts of L. Rifkin, MorphoGen Pharmaceuti-
cals, Inc., New York, NY. We thank D. Hixson for the
generous gift of his antibodies OC2, OC3, OC4, OC5,
OC10, DPP-IV, OV6, H-1, H-4, and LAP for endodermal
lineage cells and CEA-CAM1 for embryonic cells. The
following antibodies were obtained from the Developmen-
tal Studies Hybridoma Bank developed under the aus-
pices of the NICHD and maintained by the University of
Iowa, Department of Biological Sciences, Iowa City, IA:
MC480, MC631, and MC813–70 developed by D. Solter;
FORSE-1 developed by P. Patterson; RAT-401 and Rip
developed by S. Hockfield; RT-97 developed by J. Wood;
8A2 developed by V. Lemmon; SV2 developed by K.M.
Buckley; VM-1 developed by V.B. Morhenn; 151-Ig devel-
oped by A. Hubbard; 40E–C developed by A. Alvarez-
Buylla; F5D developed by W.E. Wright; MF-20 and
ALD-58 developed by D.A. Fischman; A4.74 developed by
H.M. Blau; CIIC1 developed by R. Holmdahl and K. Ru-
bin; D1–9 developed by X.-J. Ye and K. Terato; 9/30/8A4
and 12/21/1C6 developed by B. Caterson; 12C5 developed
by R.A. Asher; WV1D1 (9C5) and MP111B101 developed
by M. Solursh and A. Frazen; P2B1 and P2H3 developed
by E.A. Wayner and G. Vercellotti; P8B1 developed by
E.A. Wayner and T. LeBien; HA4c19 developed by A.
Hubbard; Hermes-1 developed by E.C. Butcher; and
H4C4, H5A5, H5C5, H5C6, and H5A4 developed by J.T.
August and J.E.K. Hildreth.
LITERATURE CITED
Ailhaud G, Grimaldi P, Negrel R. 1992. Cellular and molecular as-
pects of adipose tissue development. Annu Rev Nutr 12:207–233.
Alison MR, Poulsom R, Jeffery R, Dhillon AP, Quaglia A, Jacob J,
Novelli M, Prentice G, Williamson J, Wright NA. 2000. Hepatocytes
from non-hepatic adult stem cells. Nature 406:257.
Alvarez-Buylla A, Buskirk DR, Nottebohm F. 1987. Monoclonal anti-
body reveals radial glia in adult avian brain. J Comp Neurol 264:
159 –170.
Asher RA, Scheibe RJ, Keiser HD, Bignami A. 1995. On the existence
of a cartilage-like proteoglycan and link proteins in the central
nervous system. Glia 13:294 –308.
August JT, Hildreth JEK. 2002. In: Developmental studies hybridoma
bank product catalog, year 2002. Iowa City, IA: University of Iowa.
p 11.
Bader D, Masaki T, Fischman DA. 1982. Immunochemical analysis of
myosin heavy chain during avian myogenesis in vivo and in vitro.
J Cell Biol 95:763–770.
Banerjee A, Roach MC, Trcka P, Luduena RF. 1990. Increased micro-
tubule assembly in bovine brain tubulin lacking the type III isotype
of beta-tubulin. J Biol Chem 265:1794 –1799.
Banerjee A, Roach MC, Wall KA, Lopata MA, Cleveland DW, Lu-
duena RF. 1988. A monoclonal antibody against the type II isotype
of beta-tubulin. Preparation of isotypically altered tubulin. J Biol
Chem 263:3029 –3034.
Baudier J, Glasser N, Gerard D. 1986. Ions binding to S100 proteins.
I. Calcium- and zinc-binding properties of bovine brain S100 alpha
alpha, S100a (alpha beta), and S100b (beta beta) protein: Zn2⫹
regulates Ca2⫹ binding on S100b protein. J Biol Chem 261:8192–
8203.
Beauchamp JR, Heslop L, Yu DSW, Tajbakhsh S, Kelly RG, Wernig A,
Buckingham ME, Partridge TA, Zammit PS. 2000. Expression of
CD34 and Myf5 defines the majority of quiescent adult skeletal
muscle satellite cells. J Cell Biol 151:1221–1233.

98 YOUNG AND BLACK
Beresford JN. 1989. Osteogenic stem cells and the stromal system of
bone and marrow. Clin Orthop 240:270 –280.
Bjerknes M, Cheng H. 2002. Multipotential stem cells in adult mouse
gastric epithelium. Am J Physiol Gastrointest Liver Physiol 283:
G767–G777.
Bjornson CRR, Rietze RL, Reynolds BA, Magli MC, Vescovi AL. 1999.
Turning brain into blood: a hematopoietic fate adopted by adult
neural stem cells in vivo. Science 283:534 –537.
Bonner-Weir S, Taneja M, Weir GC, Tatarkiewicz K, Song K-H,
Sharma A, O’Neil JJ. 2000. In vitro cultivation of human islets from
expanded ductal tissue. Proc Natl Acad Sci USA 97:7999 – 8004.
Bosch P, Musgrave DS, Lee JY, Cummins J, Shuler T, Ghivizzani TC,
Evans T, Robbins TD, Huard J. 2000. Osteoprogenitor cells within
skeletal muscle. J Orthop Res 18:933–944.
Bowerman SG, Taylor SS, Putnam L, Young HE, Lucas PA. 1991.
Transforming growth factor-␤ (TGF-␤) stimulates chondrogenesis
in cultured embryonic mesenchymal cells. Surg Forum XLII:535–
536.
Brazelton TR, Rossi FM, Keshet GI, Blau HM. 2000. From marrow to
brain: expression of neuronal phenotypes in adult mice. Science
290:1775–1779.
Burgeson RE, Hollister DW. 1979. Collagen heterogeneity in human
cartilage: identification of several new collagen chains. Biochem
Biophys Res Commun 87:1124 –1131.
Butcher EC. 2002. In: Developmental studies hybridoma bank prod-
uct catalog, year 2002. Iowa City, IA: University of Iowa. p 12.
Campion DR. 1984. The muscle satellite cell: a review. Int Rev Cytol
87:225–251.
Caplan AI. 1991. Mesenchymal stem cells. J Orthop Res 9:641– 650.
Caplan AI, Elyaderani M, Mochizuki Y, Wakitani S, Goldberg V.
1997. Principles of cartilage repair and regeneration. Clin Orthop
Rel Res 342:254 –269.
Carlson BM. 1999. In: Human embryology & developmental biology,
2nd edition. St. Louis: Mosby Inc.
Caterson B, Baker JR, Christner JE, Lee Y, Lentz M. 1985. Monoclo-
nal antibodies as probes for determining the microheterogeneity of
the link proteins of cartilage proteoglycan. J Biol Chem 260:11348 –
11356.
Caterson B. 2001. In: Developmental studies hybridoma bank product
catalog, year 2001, NICHHD. Iowa City, IA: University of Iowa. p
12.
Clarke DL, Johansson CB, Wilbertz J, Veress B, Nilsson E, Karlstrom
H, Lendahl U, Frisen J. 2000. Generalized potential of adult neural
stem cells. Science 288:1660 –1663.
Colter DC, Class R, DiGirolamo CM, Prockop DJ. 2000. Rapid expan-
sion of recycling stem cells in cultures of plastic-adherent cells from
human bone marrow. Proc Natl Acad Sci USA 97:3213–3218.
Colter DC, Sekiya I, Prockop DJ. 2001. Identification of a subpopula-
tion of rapidly self-renewing and multipotential adult stem cells in
colonies of human marrow stromal cells. Proc Natl Acad Sci USA
98:7841–7845.
Cornelius JG, Tchernev V, Kao KJ, Peck AB. 1997. In vitro-generation
of islets in long-term cultures of pluripotent stem cells from adult
mouse pancreas. Horm Metab Res 29:271–277.
Cruess RL. 1982. The musculoskeletal system: embryology, biochem-
istry, and physiology. New York: Churchill Livingston. p 1–33,
109 –169, 255–287.
Damjanov I, Fox N, Knowles BB, Solter D, Lange BH, Fraley EE.
1982. Immunohistochemical localization of murine stage specific
embryonic antigens in human testicular germ cell tumors. Am J
Pathol 108:225–230.
Deasy BM, Jankowski RJ, Huard J. 2001. Muscle-derived stem cells:
characterization and potential for cell-mediated therapy. Blood
Cells Mol Dis 27:924 –933.
Deasy BM, Qu-Peterson Z, Greenberger JS, Huard J. 2002. Mecha-
nisms of muscle stem cell expansion with cytokines. Stem Cells
20:50 – 60.
Debus E, Weber K, Osborn M. 1983a. Monoclonal antibodies specific
for glial fibrillary acidic (GFA) protein and for each of the neuro-
filament triplet polypeptides. Differentiation 25:193–203.
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Debus E, Weber K, Osborn M. 1983b. Monoclonal antibodies to
desmin, the muscle-specific intermediate filament protein. EMBO J
2:2305–2312.
Dell’Agnola C, Rabascio C, Mancuso P, Capillo M, Pruneri G, Gobbi A,
Minucci S, Ronzoni S, Volorio S, Calabrese L, Tradati N, Martinelli
G, Shultz L, Bertolini F. 2002. In vitro and in vivo hematopoietic
potential of human stem cells residing in muscle tissue. Exp He-
matol 30:905–914.
Deng W, Obrocka M, Fischer I, Prockop DJ. 2001. In vitro differenti-
ation of human marrow stromal cells into early progenitors of
neural cells by conditions that increase intracellular cyclic AMP.
Biochem Biophys Res Commun 282:148 –152.
Detn CL, Latchman DS. 1993. In: Latchman DS, editor. Transcription
factors: a practical approach. New York: Oxford University Press. p
1–26.
Dittel BN, McCarthy JB, Wayner EA, LeBien TW. 1993. Regulation of
human B-cell precursor adhesion to bone marrow stromal cells by
cytokines that exert opposing effects on the expression of vascular
cell adhesion molecule-1 (VCAM-1). Blood 81:2272–2282.
Dixson K, Murphy RW, Southerland SS, Young HE, Dalton ML,
Lucas PA. 1996. Recombinant human bone morphogenetic pro-
teins-2 and 4 (rhBMP-2 and rhBMP-4) induce several mesenchymal
phenotypes in culture. Wound Rep Reg 4:374 –380.
Doetsch F, Caille I, Lim DA, Garcia-Verdugo JM, Alvarez-Buylla A.
1999. Subventricular zone astrocytes are neural stem cells in the
adult mammalian brain. Cell 97:703–716.
Donovan PJ, Gearhart J. 2001. The end of the beginning for pluripo-
tent stem cells. Nature 414:92–927.
Drazba J, Pierce M, Lemmon V. 1991. Studies of the developing chick
retina using monoclonal antibody 8A2 that recognizes a novel set of
gangliosides. Dev Biol 145:154 –163.
Eglitis MA, Mezey E. 1997. Hematopoietic cells differentiate into both
microglia and macroglia in the brains of adult mice. Proc Natl Acad
Sci USA 94:4080 – 4085.
Eisenberg CA, Markwald RR. 1997. Mixed cultures of avian blasto-
derm cells and the quail mesoderm cell line QCE-6 provide evidence
for the pluripotentiality of early mesoderm. Dev Biol 191:167–181.
Eisenberg CA, Gourdie RG, Eisenberg LM. 1997. Wnt-11 is expressed
in early avian mesoderm and required for the differentiation of the
quail mesoderm cell line QCE-6. Development 124:525–536.
Estrera VT, Luo W, Phan D, Earley K, Hixson DC. 1999. The cyto-
plasmic domain of C-CAM1 tumor suppressor is necessary and
sufficient for suppressing the tumorigenicity of prostate cancer
cells. Biochem Biophys Res Commun 263:797– 803.
Faris RA, McEntire KD, Thompson NL, Hixson DC. 1990. Identifica-
tion and characterization of a rat hepatic oncofetal membrane gly-
coprotein. Cancer Res 50:4755– 4763.
Faris RA, Monfils BA, Dunsford HA, Hixson DC. 1991. Antigenic
relationship between oval cells and a subpopulation of hepatic foci,
nodules and carcinomas induced by the ‘Resistant Hepatocyte’
model system. Cancer Res 51:1308 –1317.
Feany MB, Lee S, Edwards RH, Buckley KM. 1992. The synaptic
vesicle protein SV2 is a novel type of transmembrane transporter.
Cell 70:861– 867.
Ferrari G, Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo
A, Cossu G, Mavilio F. 1998. Muscle regeneration by bone marrow-
derived myogenic progenitors. Science 279:1528 –1530.
Forbes SJ, Vig P, Poulsom R, Wright NA, Alison MR. 2002. Adult stem
cell plasticity: new pathways of tissue regeneration become visible.
Clin Sci (Lond) 103:355–369.
Franke FE, Schachenmayr W, Osborn M, Altmannsberger M. 1991.
Unexpected immunoreactivities of intermediate filament antibodies
in human brain and brain tumors. Am J Pathol 139:67–79.
Frid MG, Shekhonin BV, Koteliansky VE, Glukhova MA. 1992. Phe-
notypic changes of human smooth muscle cells during development:
late expression of heavy caldesmon and calponin. Dev Biol 153:185–
193.
Friedman B, Hockfield S, Black JA, Woodruff KA, Waxman SG. 1989.
In situ demonstration of mature oligodendrocytes and their
processes: an immunocytochemical study with a new monoclonal
antibody, rip. Glia 2:380 –390.

ADULT STEM CELLS
Gage FH, Coates PW, Palmer TD, Kuhn HG, Fisher LJ, Suhonen JO,
Peterson DA, Suhr ST, Ray J. 1995. Survival and differentiation of
adult neuronal progenitor cells transplanted to the adult brain.
Proc Natl Acad Sci USA 92:11879 –11883.
Galli R, Borello U, Gritti A, Minasi MG, Bjornson C, Coletta M, Mora
M, De Angelis MGC, Fiocco R, Cossu G, Vescovi AL. 2000. Skeletal
myogenic potential of human and mouse neural stem cells. Nat
Neurosci 3:986 –991.
Gordon GJ, Coleman WB, Hixson DC, Grisham JW. 2000. Liver
regeneration in rats with retrorsine-induced hepatocellular injury
proceeds through a novel cellular response. Am J Pathol 156:607–
619.
Gorski JP, Griffin D, Dudley G, Stanford C, Thomas R, Huang C, Lai
E, Karr B, Solursh M. 1990. Bone acidic glycoprotein-75 is a major
synthetic product of osteoblastic cells and localized as 75- and/or
50-kDa forms in mineralized phases of bone and growth plate and
in serum. J Biol Chem 265:14956 –14963.
Grande DA, Southerland SS, Manji R, Pate DW, Schwartz RE, Lucas
PA. 1995. Repair of articular cartilage defect using mesenchymal
stem cells. Tissue Eng 1:345–353.
Gritti A, Parati EA, Cova L, Frolichsthal P, Galli R, Wanke E, Fara-
velli L, Morassutti DJ, Roisen F, Nickel DD, Vescovi AL. 1996.
Multipotential stem cells from the adult mouse brain proliferate
and self-renew in response to basic fibroblast growth factor. J Neu-
rosci 16:1091–1100.
Gritti A, Frolichsthal-Schoeller P, Galli R, Parati EA, Cova L, Pagano
SF, Bjornson CR, Vescovia AL. 1999. Epidermal and fibroblast
growth factors behave as mitogenic regulators for a single multipo-
tent stem cell-like population from the subventricular region of the
adult mouse forebrain. J Neurosci 19:3287–3297.
Grounds MD, Garrett KL, Lai MC, Wright WE, Beilharz MW. 1992.
Identification of muscle precursor cells in vivo by use of MyoD1 and
myogenin probes. Cell Tissue Res 267:99 –104.
Gui L, Chang A, Solovey M, Brown PV, Keller G, Hebbel RP. 1992.
P1H12 as an embryonic endothelial-endothelial adhesion molecule
and possibly hemangioblast marker. Blood 92(Suppl 1):697.
Gussoni E, Soneoka Y, Strickland CD, Buzney EA, Khan MK, Flint
AF, Kunkel LM, Mulligan RC. 1999. Dystrophin expression in the
mdx mouse restored by stem cell transplantation. Nature 401:390 –
394.
Hayflick L. 1965. The limited in vitro lifetime of human diploid cell
strains. Exp Cell Res 37:614 – 636.
Hixson DC, Ponce MDL, Allison JP, Walborg Jr EF. 1984. Cell surface
expression by adult rat hepatocytes of a non-collagen glycoprotein
present in rat liver biomatrix. Exp Cell Res 152:402– 414.
Hixson DC, Faris RA, Thompson NL. 1990. An antigenic portrait of
the liver during carcinogenesis. Pathobiology 58:65–77.
Hixson DC, Brown J, McBride A, Affigne S. 2000. Differentiation
status of rat ductal cells and ethionine-induced hepatic carcinomas
defined with surface-reactive monoclonal antibodies. Exp Mol
Pathol 68:152–169.
Hockfield S, McKay RD. 1985. Identification of major cell classes in
the developing mammalian nervous system. J Neurosci 5:3310 –
3328.
Holmdahl R, Rubin K, Klareskog L, Larsson E, Wigzell H. 1986.
Characterization of the antibody response in mice with type II
collagen-induced arthritis, using monoclonal anti-type II collagen
antibodies. Arthritis Rheum 29:400 – 410.
Hubbard AL, Bartles JR, Braiterman LT. 1985. Identification of rat
hepatocyte plasma membrane proteins using monoclonal antibod-
ies. J Cell Biol 100:1115–1125.
Jackson KA, Mi T, Goodell MA. 1999. Hematopoietic potential of stem
cells isolated from murine skeletal muscle. Proc Natl Acad Sci USA
96:14482–14486.
Jackson KA, Majka SM, Wang H, Pocius J, Hartley CJ, Majesky MW,
Entman ML, Michael LH, Hirschi KK, Goodell MA. 2001. Regener-
ation of ischemic cardiac muscle and vascular endothelium by adult
stem cells. J Clin Invest 107:1395–1402.
Janes SM, Lowell S, Hutter C. 2002. Epidermal stem cells. J Pathol
197:479 – 491.
Jankowski RJ, Haluszczak C, Trucco M, Huard J. 2001. Flow cyto-
metric characterization of myogenic cell populations obtained via
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
99
the preplate technique: potential for rapid isolation of muscle-de-
rived stem cells. Hum Gene Ther 12:619 – 628.
Javazon EH, Colter DC, Schwarz EJ, Prockop DJ. 2001. Rat marrow
stromal cells are more sensitive to plating density and expand more
rapidly from single-cell-derived colonies than human marrow stro-
mal cells. Stem Cells 19:219 –225.
Jiang Y, Vaessen B, Lenvik T, Blackstad M, Reyes M, Verfaillie CM.
2002. Multipotent progenitor cells can be isolated from bone mar-
row, muscle and brain. Exp Hematol 30:896 –904.
Johansson CB, Momma S, Clarke DL, Risling M, Lendahl U, Frisen J.
1999. Identification of a neural stem cell in the adult mammalian
central nervous system. Cell 96:25–34.
Johe KK. 2003. Development of neural stem cells for treatment of
neurological diseases. Arch Neurol 60:297.
Joshi HC, Cleveland DW. 1990. Diversity among tubulin subunits:
toward what functional end? Cell Motil Cytoskel 16:159.
Kacsoh B. 2000. Endocrine physiology. New York: McGraw-Hill.
Kasugai S, Nagata T, Sodek J. 1992. Temporal studies on the tissue
compartmentalization of bone sialoprotein (BSP), osteopontin
(OPN), and SPARC protein during bone formation in vitro. J Cell
Physiol 152:467– 477.
Kishimoto T, Kikutani H, Borne AEGKrvd, Goyert SM, Mason D,
Miyasaka M, Moretta L, Okumura K, Shaw S, Springer T, Sug-
amura K, Zola H. 1997. Leucocyte typing VI, white cell differenti-
ation antigens. Hamden, CT: Garland Publishing.
Kopen GC, Prockop DJ, Phinney DG. 1999. Marrow stromal cells
migrate throughout forebrain and cerebellum, and they differenti-
ate into astrocytes after injection into neonatal mouse brains. Proc
Natl Acad Sci USA 96:10711–10716.
Krause DS, Theise ND, Collector Mi, Henegariu O, Hwang S, Gardner
R, Neutzel S, Sharkis SJ. 2001. Multi-organ, multi-lineage engraft-
ment by a single bone marrow derived stem cell. Cell 105:369 –377.
Krebsbach PH, Robey PG. 2002. Dental and skeletal stem cells: po-
tential cellular therapeutics for craniofacial regeneration. J Dental
Educ 66:766 –773.
Kruger GM, Mosher JT, Bixby S, Joseph N, Iwashita T, Morrison SJ.
2002. Neural crest stem cells persist in the adult gut but undergo
changes in self-renewal, neuronal subtype potential, and factor
responsiveness. Neuron 35:657– 669.
Kumagai J, Sarkar K, Uhthoff HK, Okawara Y, Ooshima A. 1994.
Immunohistochemical distribution of type I, II and III collagens in
the rabbit supraspinatus tendon insertion. J Anat 185:279 –284.
Kuznetsov SA, Mankani MH, Gronthos S, Satomura K, Bianco P,
Robey PG. 2001. Circulating skeletal stem cells. J Cell Biol 153:
1133–1139.
LaBarge MA, Blau HM. 2002. Biological progression from adult bone
marrow to mononucleated muscle stem cell to multinucleate muscle
fiber in response to injury. Cell 111:589 – 601.
Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, Osborne
L, Wang X, Finegold M, Weisman IL, Grompe M. 2000. Purified
hematopoietic stem cells can differentiate into hepatocytes in vivo.
Nat Med 6:1229 –1234.
Lannagi R, Cochran NA, Ishigami F, Hakomori S-I, Andrews PW,
Knowles BB, Solter D. 1983. Stage specific embryonic antigens
(SSEA-3 and -4) are epitopes of a unique globo-series ganglioside
isolated from human treatocarcinoma cells. EMBO J 2:2355–2361.
Lazard D, Sastre X, Frid MG, Glukhova MA, Thiery JP, Koteliansky
VE. 1993. Expression of smooth muscle-specific proteins in myoepi-
thelium and stromal myofibroblasts of normal and malignant hu-
man breast tissue. Proc Natl Acad Sci USA 90:999 –1003.
Lee JY, Qu-Petersen Z, Cao B, Kimura S, Jankowski R, Cummins J,
Usas A, Gates C, Robbins P, Wernig A, Huard J. 2000. Clonal
isolation of muscle-derived cells capable of enhancing muscle regen-
eration and bone healing. J Cell Biol 150:1085–1099.
Lie DC, Dziewczapolski G, Willhoite AR, Kaspar BK, Shults CW,
Gage FH. 2002. The adult substantia nigra contains progenitor cells
with neurogenic potential. J Neurosci 22:6639 – 6649.
Lucas PA, Syftestad GT, Caplan AI. 1986. Partial isolation and char-
acterization of a chemotactic factor from adult bovine bone for
mesenchymal cells. Bone 7:365–371.

100 YOUNG AND BLACK
Lucas PA, Caplan AI. 1988. Chemotactic response of embryonic limb
bud mesenchymal cells and muscle-derived fibroblasts to trans-
forming growth factor-beta. Connect Tissue Res 18:1–7.
Lucas PA, Price PA, Caplan AI. 1988. Chemotactic response of mes-
enchymal cells, fibroblasts and osteoblast-like cells to bone Gla
protein. Bone 5:19 –23.
Lucas PA. 1989. Chemotactic response of osteoblast-like cells to trans-
forming growth factor-beta. Bone 10:459 – 463.
Lucas PA, Calcutt AF, Mulvaney DJ, Young HE, Southerland SS.
1992. Isolation of putative mesenchymal stem cells from rat embry-
onic and adult skeletal muscle. In Vitro Cell Dev Biol 28:154A.
Lucas PA, Calcutt AF, Ossi P, Young HE, Southerland SS. 1993.
Mesenchymal stem cells from granulation tissue. J Cell Biochem
17E:122.
Lucas PA, Young HE, Laurencin CT. 1994a. Muscle morphogenetic
protein and use thereof. U.S. Patent 5,328,695.
Lucas PA, Calcutt AF, Ossi P, Young HE, Southerland SS. 1994b.
Granulation tissue contains cells capable of differentiating into
multiple mesenchymal phenotypes. J Cell Biochem 18C:276.
Lucas PA, Calcutt AF, Southerland SS, Wilson JA, Harvey RL, Ware-
jcka DJ, Young HE. 1995. A population of cells resident within
embryonic and newborn rat skeletal muscle is capable of differen-
tiating into multiple mesodermal phenotypes. Wound Rep Reg
3:457– 468.
Majka SM, Jackson KA, Kienstra KA, Majesky MW, Goodell MA,
Hirschi KK. 2003. Distinct progenitor populations in skeletal mus-
cle are bone marrow derived and exhibit different cell fates during
vascular regeneration. J Clin Invest 111:71–79.
Mauro A. 1961. Satellite cell of skeletal muscle fibers. J Biophys
Biochem Cytol 9:493– 498.
McGuire WP. 1998. High-dose chemotherapy and autologous bone
marrow or stem cell reconstitution for solid tumors. Curr Probl
Cancer 22:135–137.
McKinney-Freeman SL, Jackson KA, Camargo FD, Ferrari G, Mavil-
lio F, Goodell MA. 2002. Muscle-derived hematopoietic stem cells
are hematopoietic in origin. Proc Natl Acad Sci USA 99:1341–1346.
Mezey E, Chandross KJ, Harta G, Maki RA, McKercher SR. 2000.
Turning blood into brain: cells bearing neuronal antigens generated
in vivo from bone marrow. Science 290:1779 –1782.
Moore KL, Persaud TVN. 1998. The developing human: clinically
oriented embryology, 6th edition. Philadelphia: W.B. Saunders Co.
Morhenn VB. 2002. In: Developmental studies hybridoma bank prod-
uct catalog, year 2002. Iowa City, IA: University of Iowa. p 24.
Mujoo K, Ali M, Sahib MK. 1983. Isolation, characterization, and
synthesis of alpha-fetoprotein from neonatal rat brain. J Neuro-
chem 41:1223–1228.
Muraglia A, Cancedda R, Quarto R. 2000. Clonal mesenchymal pro-
genitors from bone marrow differentiate in vitro according to a
hierarchial model. J Cell Sci 113:1161–1166.
Murayama A, Matsuzaki Y, Kawaguchi A, Shimazaki T, Okano H.
2002. Flow cytometric analysis of neural stem cells in the develop-
ing and adult mouse brain. J Neurosci Res 69:837– 847.
Naumann K, Pette D. 1994. Effects of chronic stimulation with dif-
ferent impulse patterns on the expression of myosin isoforms in rat
myotube cultures. Differentiation 55:203–211.
Oben JA, Yang S, Lin H, Ono M, Diehl AM. 2003. Acetylcholine
promotes the proliferation and collagen gene expression of myofi-
broblastic hepatic stellate cells. Biochem Biophys Res Commun
300:172–177.
Otani A, Kinder K, Ewalt K, Otero FJ, Schimmel P, Friedlander M.
2002. Bone marrow-derived stem cells target retinal astrocytes and
can promote or inhibit retinal angiogenesis. Nat Med 8:1004 –1010.
Owen M. 1988. Marrow stromal cells. J Cell Sci Suppl 10:63–76.
Owen M, Friedenstein AJ. 1988. Stromal stem cells: marrow-derived
osteogenic precursors. Ciba Found Symp 136:42– 60.
Palis J, Segel GB. 1998. Developmental biology of erythropoiesis.
Blood Rev 12:1061–1064.
Palmer TD, Schwartz PH, Taupin P, Kaspar B, Stein SA, Gage FH.
2001. Progenitor cells from human brain after death. Nature 411:
42– 43.
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Pate DW, Southerland SS, Grande DA, Young HE, Lucas PA. 1993.
Isolation and differentiation of mesenchymal stem cells from rabbit
muscle. Surg Forum XLIV:587–589.
Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Mu-
rase N, Boggs SS, Greenberger JS, Goff JP. 1999. Bone marrow as
a potential source of hepatic oval cells. Science 284:1168 –1170.
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca
JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. 1999.
Multilineage potential of adult human mesenchymal stem cells.
Science 148:143–147.
Poulsom R, Alison MR, Forbes SJ, Wright NA. 2002. Adult stem cell
plasticity. J Pathol 197:441– 456.
Prockop DJ. 1997. Marrow stromal cells as stem cells for nonhema-
topoietic tissues. Science 276:71–74.
Quirici N, Soligo D, Bossolasco P, Servida F, Sumini C, Deliliers GL.
2002. Isolation of bone marrow mesenchymal stem cells by anti-
nerve growth factor receptor antibodies. Exp Hematol 30:783–791.
Ramiya VK, Maraist M, Arfos KE, Schatz DA, Peck AB, Cornelius JG.
2000. Reversal of insulin-dependent diabetes using islets generated
in vitro from pancreatic stem cells. Nat Med 6:278 –282.
Ratajczak MZ, Pletcher CH, Marlicz W, Machlinski B, Moore J, Wasik
M, Ratajczak J, Gewirtz AM. 1998. CD34⫹, kit⫹, rhodamine 123
(low) phenotype identifies a marrow cell population highly enriched
for human hematopoietic stem cells. Leukemia 12:942–950.
Reyes M, Verfaillie CM. 2001. Characterization of multipotent adult
progenitor cells, a subpopulation of mesenchymal stem cells. Ann
NY Acad Sci 938:231–233; discussion, 233–235.
Reyes M, Lund T, Lenvik T, Aguiar D, Koodle L, Verfaillie CM. 2001.
Purification and ex vivo expansion of postnatal human marrow
mesodermal progenitor cells. Blood 98:2615–2625.
Reynolds R, Carey EM, Herschkowitz N. 1989. Immunohistochemical
localization of myelin basic protein and 2⬘,3⬘-cyclicnucleotide 3⬘-
phosphohydrolase in flattened membrane expansions produced by
cultured oligodendrocytes. Neuroscience 28:181–188.
Rogers JJ, Young HE, Adkison LR, Lucas PA, Black Jr AC. 1995.
Differentiation factors induce expression of muscle, fat, cartilage,
and bone in a clone of mouse pluripotent mesenchymal stem cells.
Am Surg 61:231–236.
Romero-Ramos M, Vourc’h P, Young HE, Lucas PA, Wu Y, Chi-
vatakarn O, Zaman R, Dunkelman N, El-Kalay MA, Chesselet M-F.
2002. Neuronal differentiation of stem cells isolated from adult
muscle. J Neurosci Res 69:894 –907.
Ronnov-Jessen L, Celis JE, Van Deurs B, Petersen OW. 1992. A
fibroblast-associated antigen: characterization in fibroblasts and
immunoreactivity in smooth muscle differentiated stromal cells.
J Histochem Cytochem 40:475– 486.
Saito T, Dennis JE, Lennon DP, Young RG, Caplan AI. 1995. Myo-
genic expression of mesenchymal stem cells within myotubes of
mdx mice in vitro and in vivo. Tissue Eng 1:327–343.
Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T,
Willing A, Freeman TB, Saporta S, Janssen W, Patel N, Cooper DR,
Sanberg PR. 2000. Adult bone marrow stromal cells differentiate
into neural cells in vitro. Exp Neurol 164:247–256.
Schaart G, Moens L, Endert JM, Ramaekers FCS. 1997. Biochemical
characterization of cardiotin, a sarcoplasmic reticulum associated
protein. FEBS Lett 403:168 –172.
Shafiq SA, Shimizu T, Fischman DA. 1984. Heterogeneity of type I
skeletal muscle fibers revealed by monoclonal antibody to slow
myosin. Muscle Nerve 7:380 –387.
Shoptaw JH, Bowerman S, Young HE, Lucas PA. 1991. Use of gelfoam
as a substrate for osteogenic cells of marrow. Surg Forum XLII:537–
538.
Simper D, Stalboerger PG, Panetta CJ, Wang S, Caplice NM. 2002.
Smooth muscle progenitor cells in human blood. Circulation 106:
1199 –1204.
Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillessen D, Gabbiani
G A. 1986. A monoclonal antibody against alpha-smooth muscle
actin: a new probe for smooth muscle differentiation. J Cell Biol
103:2787–2796.
Solovey A, Lin Y, Browne P, Choong S, Wayner E, Hebbel RP. 1997.
Circulating activated endothelial cells in sickle cell anemia. N Eng
J Med 337:1584 –1590.

ADULT STEM CELLS
Solter D, Knowles BB, 1978. Monoclonal antibody defining a stage-
specific mouse embryonic antigen (SSEA-1). Proc Natl Acad Sci
USA 75:5565–5569.
Sprinkle TJ, Agee JF, Tippins RB, Chamberlain CR, Faguet GB,
DeVries GH. 1987. Monoclonal antibody production to human and
bovine 2⬘,3⬘-cyclic nucleotide 3⬘-phosphodiesterase (CNPase): high-
specificity recognition in whole brain acetone powders and conser-
vation of sequence between CNP1 and CNP2. Brain Res 426:349 –
357.
Sprinkle T. 1989. 2⬘,3⬘-cyclic nucleotide 3⬘-phosphodiesterase, an oli-
godendrocyte-Schwann cell and myelin-associated enzyme of the
nervous system. CRC Crit Rev Neurobiol 4:235–301.
St. Croix B, Rago C, Velculescu V, Traverso G, Romans KE, Mont-
gomery E, Lal A, Riggins GJ, Lengauer C, Vogelstein B, Kinzler
KW. 2000. Genes expressed in human tumor endothelium. Science
289:1197–1202.
Tanguchi H, Toyoshima T, Fukao K, Nakauchi H. 1996. Presence of
hematopoietic stem cells in the adult liver. Nat Med 2:198 –203.
Taylor DA, Atkins BZ, Hungspreugs P, Jones TR, Reedy MC, Hutch-
enson KA, Glower DD, Kraus WE. 1997. Delivery of primary autol-
ogous skeletal myoblasts into rabbit heart by coronary infusion: a
potential approach to myocardial repair. Proc Assoc Am Physicians
109:245–253.
Thulasi R, Dias P, Houghton PJ, Houghton JA. 1996. Alpha 2a-
interferon-induced differentiation of human alveolar rhabdomyo-
sarcoma cells: correlation with down-regulation of the insulin-like
growth factor type I receptor. Cell Growth Differ 7:531–541.
Tole S, Kaprielian Z, Ou SK, Patterson PH. 1995. FORSE-1: a posi-
tionally regulated epitope in the developing rat central nervous
system. J Neurosci 15:957–969.
Tole S, Patterson PH. 1995. Regionalization of the developing
forebrain: a comparison of FORSE-1, Dlx-2, and BF-1. J Neurosci
15:970 –980.
Toma JG, Akhavan M, Fernandes KJ, Barnabe-Heider F, Sadikot A,
Kaplan DR, Miller FD. 2001. Isolation of multipotent stem cells
from the dermis of mammalian skin. Nat Cell Biol 3:778 –784.
Tomita S, Li RK, Weisel RD, Mickle DA, Kim EJ, Sakai T, Jia ZQ.
1999. Autologous transplantation of bone marrow cells improves
damaged heart function. Circulation 100(Suppl II):11247–11256.
Tsai RY, Kittappa R, McKay RGD. 2002. Plasticity, niches, and the
use of stem cells. Dev Cell 2:707–712.
Turgeman G, Pittman DD, Muller R, Kurkalli BG, Zhou S, Pelled G,
Peyser A, Zilberman Y, Moutsatsos IK, Gazit D. 2001. Engineered
human mesenchymal stem cells: a novel platform for skeletal cell
mediated gene therapy. J Gene Med 3:240 –251.
Vacanti MP, Leonard JL, Dore B, Bonassar LJ, Cao Y, Stachelek SJ,
Vacaqnti JP, O’Connell F, Yu CS, Farwell AP, Vacanti CA. 2001a.
Tissue-engineered spinal cord. Transplant Proc 33:592–598.
Vacanti MP, Roy A, Cortiella J, Bonassar L, Vacanti CA. 2001b.
Identification and initial characterization of spore-like cells in adult
mammals. J Cell Biochem 80:455– 460.
Vescovi AL, Galli R, Gritti A. 2001. The neural stem cells and their
transdifferentiation capacity. Biomed Pharmacother 55:201–205.
Vierck JL, McNamara JP, Dodson MV. 1996. Proliferation and differ-
entiation of progeny of ovine unilocular fat cells (adipofibroblasts).
In Vitro Cell Dev Biol Anim 32:564 –572.
Walborg Jr EF, Tsuchida S, Weeden DS, Thomas MW, Barrick A,
McEntire KD, Allison JP, Hixson DC. 1985. Identification of dipep-
tidyl peptidase IV as a protein shared by the plasma membrane of
hepatocytes and liver biomatrix. Exp Cell Res 158:509 –518.
Warejcka DJ, Harvey R, Taylor BJ, Young HE, Lucas PA. 1996. A
population of cells isolated from rat heart capable of differentiating
into several mesodermal phenotypes. J Surg Res 62:233–242.
Webster C, Silberstein L, Hays AP, Blau HM. 1988. Fast muscle fibers
are preferentially affected in Duchenne muscular dystrophy. Cell
52:503–513.
Winter A, Breit S, Parsch D, Benz K, Steck E, Hauner H, Weber RM,
Ewerbeck V, Richter W. 2003. Cartilage-like gene expression in
differentiated human stem cell spheroids. Arthritis Rheum 48:418 –
429.
Wood JN, Anderton BH. 1981. Monoclonal antibodies to mammalian
neurofilaments. Biosci Rep 1:263–268.
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
101
Woodbury D, Schwarz EJ, Prockop DJ, Black IB. 2000. Adult rat and
human bone marrow stromal cells differentiate into neurons.
J Neurosci Res 61:364 –370.
Woodbury D, Reynolds K, Black IB. 2002. Adult bone marrow stromal
stem cells express germline, ectodermal, endodermal, and mesoder-
mal genes prior to neurogenesis. J Neurosci Res 96:908 –917.
Wright WE, Binder M, Funk W. 1991. Cyclic amplification and selec-
tion of targets (CASTing) for the myogenin consensus binding site.
Mol Cell Biol 11:4104 – 4110.
Yang L, Shiwu L, Hatch H, Ahrens K, Cornelius JG, Petersen BE,
Peck AB. 2002. In vitro trans-differentation of adult hepatic stem
cells into pancreatic endocrine hormone-producing cells. Proc Natl
Acad Sci USA 99:8078 – 8083.
Ye XJ, Terato K, Nakatani H, Cremer MA, Yoo TJ. 1991. Monoclonal
antibodies against bovine type IX collagen (LMW fragment): pro-
duction, characterization, and use for immunohistochemical local-
ization studies. J Histochem Cytochem 39:265–271.
Young HE. 1977a. Epidermal ridge formation during limb regenera-
tion in the adult salamander, Ambystoma annulatum. Proc Ark
Acad Sci 31:107–109.
Young HE. 1977b. Limb regeneration in the adult salamander, Am-
bystoma annulatum Cope 1889 (Amphibia: Ambystomatidae). Fay-
etteville, AR: University of Arkansas Library Press.
Young HE. 1977c. Anomalies during limb regeneration in the adult
salamander, Ambystoma annulatum. Proc Ark Acad Sci 31:110 –
111.
Young HE. 1983. A temporal examination of glycoconjugates during
the initiation phase of limb regeneration in adult Ambystoma.
Lubbock, TX: Texas Tech University Library Press.
Young HE, Bailey CF. Dalley BK. 1983a. Environmental conditions
prerequisite for complete limb regeneration in the postmetamorphic
adult land-phase salamander, Ambystoma. Anat Rec 206:289 –294.
Young HE, Bailey CF, Dalley BK. 1983b. Gross morphological anal-
ysis of limb regeneration in postmetamorphic adult Ambystoma.
Anat Rec 206:295–306.
Young HE, Dalley BK, Markwald RR. 1983c. Identification of hyal-
uronate within peripheral nervous tissue matrices during limb re-
generation. In: Coates PW, Markwald RR, Kenny AD, editors. De-
veloping and regenerating vertebrate nervous systems, neurology
and neurobiology. New York: Alan R. Liss, Inc. 6, p 175–183.
Young HE, Dalley BK, Markwald RR. 1983d. The interaction of gly-
cosaminoglycans (GAG) and nervous tissue regeneration in adult
Ambystoma. Anat Rec 205:202.
Young HE, Bailey CF, Markwald RR, Dalley BK. 1985. Histological
analysis of limb regeneration in postmetamorphic adult Ambys-
toma. Anat Rec 212:183–194.
Young HE, Dalley BK, Markwald RR. 1989a. Glycoconjugates in
normal wound tissue matrices during the initiation phase of limb
regeneration in adult Ambystoma. Anat Rec 223:231–241.
Young HE, Dalley BK, Markwald RR. 1989b. Effect of selected den-
ervations on glycoconjugate composition and tissue morphology
during the initiation phase of limb regeneration in adult Ambys-
toma. Anat Rec 223:231–241.
Young HE, Carrino DA, Caplan AI. 1989c. Histochemical analysis of
newly synthesized and resident sulfated glycosaminoglycans during
musculogenesis in the embryonic chick leg. J Morph 201:85–103.
Young HE, Morrison DC, Martin JD, Lucas PA. 1991. Cryopreserva-
tion of embryonic chick myogenic lineage-committed stem cells. J
Tissue Cult Methods 13:275–284.
Young HE, Ceballos EM, Smith JC, Lucas PA, Morrison DC. 1992a.
Isolation of embryonic chick myosatellite and pluripotent mesen-
chymal stem cells. J Tissue Cult Methods 14:85–92.
Young HE, Sippel J, Putnam LS, Lucas PA, Morrison DC. 1992b.
Enzyme-linked immuno-culture assay. J Tissue Cult Methods 14:
31–36.
Young HE, Ceballos EM, Smith JC, Mancini ML, Wright RP, Ragan
BL, Bushell I, Lucas PA. 1993. Pluripotent mesenchymal stem cells
reside within avian connective tissue matrices. In Vitro Cell Dev
Biol Anim 29A:723–736.
Young HE, Mancini ML, Wright RP, Smith JC, Black Jr AC, Reagan
CR, Lucas PA. 1995. Mesenchymal stem cells reside within the
connective tissues of many organs. Dev Dyn 202:137–144.

102 YOUNG AND BLACK
Young HE, Lucas PA. 1998. Pluripotent mesenchymal stem cells and
methods of use thereof. U.S. Patent 5,827,735.
Young HE, Wright RP, Mancini ML, Lucas PA, Reagan CR. Black Jr
AC. 1998a. Bioactive factors affect proliferation and phenotypic
expression in pluripotent and progenitor mesenchymal stem cells.
Wound Rep Reg 6:65–75.
Young HE, Rogers JJ, Adkison LR, Lucas PA, Black Jr AC. 1998b.
Muscle morphogenetic protein induces myogenic gene expression in
Swiss-3T3 cells. Wound Rep Reg 6:530 –541.
Young HE, Steele TA, Bray RA, Detmer K, Blake LW, Lucas PA,
Black Jr AC. 1999. Human pluripotent and progenitor cells display
cell surface cluster differentiation markers CD10, CD13, CD56, and
MHC Class-I. Proc Soc Exp Biol Med 221:63–71.
Young HE. 2000. Stem cells and tissue engineering. In: Huard J, Fu
FH, editors. Gene Therapy in orthopaedic and sports medicine. New
York: Springer-Verlag. p 143–173.
Young HE, Duplaa C, Young TM, Floyd JA, Reeves ML, Davis KH,
Mancini GJ, Eaton ME, Hill JD, Thomas K, Austin T, Edwards C,
Cuzzourt J, Parikh A, Groom J, Hudson J, Black Jr AC. 2001a.
Clonogenic analysis reveals reserve stem cells in postnatal mam-
mals. I. Pluripotent mesenchymal stem cells. Anat Rec 263:350 –
360.
Young HE, Steele T, Bray RA, Hudson J, Floyd JA, Hawkins KC,
Thomas K, Austin T, Edwards C, Cuzzourt J, Duenzl M, Lucas PA,
Black Jr AC. 2001b. Human reserve pluripotent mesenchymal stem
cells are present in the connective tissues of skeletal muscle and
dermis derived from fetal, adult, and geriatric donors. Anat Rec
264:51– 62.
Young HE. 2004. Existence of reserve quiescent stem cells in adults,
from amphibians to humans. Curr Top Microbiol Immunol 280:71–
109.
Young HE, Duplaa C, Romero-Ramos M, Chesselet M-F, Vourc’h P,
Yost MJ, Ericson K, Terracio L, Asahara T, Masuda H, Tamura-
15524892, 2004, 1, Downloaded from https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/ar.a.10134 by Cochrane Romania, Wiley Online Library on [31/10/2022]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
Ninomiya S, Detmer K, Bray RA, Steele TA, Hixson D, El-Kalay M,
Tobin BW, Russ RD, Horst MN, Floyd JA, Henson NL, Hawkins
KC, Groom J, Parikh A, Blake L, Bland LJ, Thompson AJ, Kirincich
A, Moreau C, Hudson J, Bowyer III FP, Lin TJ, Black Jr AC. 2004a.
Adult reserve stem cells and their potential for tissue engineering.
J Cell Biochem Biophys 40(1):(in press).
Young HE, Duplaa C, Yost MJ, Henson NL, Floyd JA, Detmer K,
Thompson AJ, Powell SW, Gamblin TC, Kizziah K, Holland BJ,
Boev A, Van de Water JM, Godbee DC, Jackson S, Rimando M,
Edwards CR, Wu E, Cawley C, Edwards PD, Macgregor A, Bozof R,
Thompson TM, Petro Jr GJ, Shelton HM, McCampbell BL, Mills
JC, Flynt FL, Steele TA, Kearney M, Kirincich-Greathead A, Hardy
W, Young PR, Amin AV, Williams RS, Horton MM, McGuinn S,
Hawkins KC, Davis JG, Parikh A, Ericson K, Terracio L, Moreau C,
Hixson D, Tobin BW, Hudson J, Bowyer III FP, Black Jr AC. 2004b.
Clonogenic analysis reveals reserve stem cells in postnatal mam-
mals. II. Pluripotent epiblastic-like stem cells. Anat Rec (in press).
Zhao Y, Glesne D, Huberman E. 2003. A human peripheral blood
monocyte-derived subset acts as pluripotent stem cells. Proc Natl
Acad Sci USA 100:2426 –2431.
Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim
P, Lorenz P, Hedrick MH. 2001. Multilineage cells from human
adipose tissue: implications for cell-based therapies. Tissue Eng
7:211–228.
Zulewski H, Abraham EJ, Gerlach MJ, Daniel PB, Moritz W, Muller
B, Vallejo M, Thomas MK, Habener JF. 2001. Multipotential nes-
tin-positive stem cells isolated from adult pancreatic islets differ-
entiate ex vivo into pancreatic endocrine, exocrine, and hepatic
phenotypes. Diabetes 50:521–533.
Zvaifler NJ, Marinova-Mutafchieva L, Adams G, Edwards CJ, Moss J,
Burger JA, Maini RN. 2000. Mesenchymal precursor cells in the
blood of normal individuals. Arthritis Res 2:477– 488.