Hematopoiesis o Major hematopoietic organ

of early and midfetal life

 Key Role
o Gives rise to fetal Hgb (Hgb
 RBC carries oxygen
F)
 The process of blood cell
o In the middle part of fetal
production, differentiation, and
life, the spleen and, to a
development
lesser extent, lymph nodes
Types of Human Stem Cells have minor role in
hematopoiesis
 Totipotential stem cell
o Can develop into human cell Medullary Phase
type. CORE/first one  Bone marrow
 Pluripotential stem cells o Begins in the 4th month of
o Present several days after
gestation (20 weeks)
fertilization. Develop any o After the fifth fetal month the
cell type except fetus.
primary site of
Limited drainage
hematopoiesis
 Multipotential stem cells o Until post-natal life
o Found in adults. Example in
(continuous)
bone marrow o Last phase
Early Development of Blood Cells
Bone Marrow
 Embryonic blood cells –
 Found within the cavities of all
MESODERM
bones and may be present in two
 Aorta-gonad-mesonephros region forms:
Mesoblastic Phase o Yellow marrow
 Which is normally
 Yolk Sac (fetal stage) inactive and
o Site of primitive composed mostly of
hematopoiesis fat (adipose) tissue
o Blood islands - divided o Red marrow
o Contain predominantly  Which is normally
primitive erythroblasts active in the
o Formed during the first 2 to production of most
8 weeks of life types of leukocytes,
o Produce hemoglobin (Gower- erythrocytes, and
1, Gower-2, and Portland) thrombocytes
 One of the body’s largest organs
Hepatic Phase
 Represents approximately 3.5% to
 Liver 6% of total body weight
o Sixth week (2nd – 5th month)  Averages around 1,500 g in adults,
  The hematopoietic marrow is o Function in phagocytosis, and
organized around the bone both macrophages and
vasculature (Haversian) lymphocytes secrete various
 First few years of life – red and cytokines that regulate
cellular hematopoiesis
o They are located throughout
the marrow space
 Osteoblasts
o Bone-forming cells
Extramedullary Hematopoiesis
 Osteoclasts
 Abnormal circumstances, the o Bone-resorbing cells
spleen, liver, and lymph nodes  Fibroblasts
revert back to producing  Reticular adventitial cells
immature blood cells o Form an incomplete layer of
 When the bone marrow becomes cells on the abluminal
dysfunctional in cases such as surface of the vascular
aplastic anemia, infiltration by sinuses
malignant cells, or over proliferation
of a cell line (e.g., leukemia).
 When the bone marrow is unable to
meet the demands placed on it, as in
the hemolytic anemia
Hematopoietic Microenvironment
 Endothelial cells
o Broad, flat cells that form a Lineage Specific Hematopoiesis
single continuous later along
the inner surface of the  Erythropoiesis
arteries, veins, and vascular o Occurs in distinct anatomical
sinuses. sites called erythropoietic
o Regulate the flow of islands, specialized niches in
particles entering and which erythroid precursors
leaving hematopoietic proliferate, differentiate,
spaces in the vascular sinuses and enucleate.
 Adipocytes  Granulopoiesis
o Are large cells with a single o Can be recognized as a
fat vacuole maturational unit. Maturing
o They play a role in cells spend an average of 3
to 6 days in the
regulating the volume of the
proliferating pool. Average
marrow in which active
life span of 6 to 10 hours in
hematopoiesis occurs
circulation
 Macrophages
  Lymphocytes and plasma cells are o Chromosome 7
produced in lymphoid follicles  GM-CSF, IL-3, and M-CSF
 Megakaryopoiesis o Long arm of chromosome 5
o Takes place adjacent to the  G-CSF
sinus endothelium. Develop o Chromosome 17
into platelets in
approximately 5 days.
Interleukins Characteristics of Human
Hematopoietic Growth Factors
 Cytokines that act independently or Growth Cellular Progen Mature
in conjunction with other Factor Source itor Cell
interleukins to encourage Cell Target
hematopoietic growth Target
 Used as signaling molecules in many Erythrop Peritubu CFU- None
cells of the body oietin lar cells E, late
of the BFU-
Abbreviated List of Cytokines and kidney, E,
Growth Factors Kupffer CFU-
Cytokine Cell Modifier cells Meg
IL-2 T cells, B cells, NK IL-3 Activate CFU- Eosinop
cells d T blast, hils,
IL-3 Multilineage lymphoc CFU- monocyt
stimulating factor ytes GEM es
IL-4 B cells, T cells, M,
mast cells CFU-
IL-6 Stem cells, B cells GM,
IL-7 Pre-B cells, T cells, CFU-
early granulocytes G,
IL-11 Megakaryocytes CFU-
GM-CSF Granulocytes, M,
macrophages, CFU-
fibroblasts, Eo,
endothelial cells CFU-
EPO Red cell progenitor Meg,
cells CFU-
Baso,
BFU-E
Hematopoietic Growth Factor G-CSF Monocy CFU-G Granulo
tes, cytes
 Regulating the proliferation and fibrobla
differentiation of HPCs as well as sts,
regulating the survival and function endothel
of mature blood cells ial cells
 Encoded by a single gene M-CSF Monocy CFU- Monocy
 Erythropoietin tes, M tes
 fibrobla  Granulation
sts, o None – non-specific –
endothel specific
ial cells  Shape
GM-CSF T CFU- Granulo
o Most distinctive variation in
lymphoc blast, cytes
ytes, CFU- cytoplasmic shape occurs in
monocyt GEM some blast forms,
es, M, monocytes, and
eosinop CFU- megakaryocytes.
hils, GM,
monocyt CFU-  Quantity of cytoplasm
es, G, o Cytoplasm increases as age
fibrobla CFU- increases
sts, Eo,
 Vacuolization
endothel CFU-
o Vacuoles increases as age
ial cells Meg,
BFU-E increases
Examination of Maturing Blood Cells o Artifacts produce of the
General Cellular Characteristics blood is stored for a longer-
 Overall cell size than-acceptable period
o Increase Size as  Inclusion bodies
AGE/MATURITY decreases
 Nuclear-cytoplasmic ratio
o Increases NUCLEAR SIZE
decreases as
AGE/MATURITY, NC ratio
decreases as
AGE/MATURITY increases
General Nuclear Characteristics
 Chromatin pattern
o Loose-Clumped
 Nuclear shape
o Very distinctive for particular
cell types
 Presence of nucleoli (blast)
o As cells mature, nucleoli are
usually not visible
Cytoplasmic Characteristics
 Staining color and intensity
o Dark blue – lighter blue
 atophilic) atophilic) atophilic)
erythrocyte erythrocyte erythrocyte
Erythrocyte Erythrocyte Erythrocyte

Pronormoblast/Rubriblast/
Red Blood Cells
Proerythroblast
Terminologies
 RBCs are formally called  Largest of the erythroid
erythrocytes precursors
 Nucleated precursors in the bone  The “mother cell”
marrow are called erythroblasts  Nucleus
 Developing nucleated cells with o N:C ratio: 4:1
normal appearance can be called o Chromatin: fine and dark
normoblasts staining
 Rubriblast terminology parallels the o Nucleoli: 1- 2
nomenclature used for granulocyte  Cytoplasm:
development. o Heterogenous
 Nuclei are always “baseball” round o Dark blue (distinctively
 As hemoglobin develops, the basophilic)
cytoplasm becomes more magenta. o No granules
Normoblast Rubriblasti Erythrobla Prorubricyte/ Basophilic Normoblast
ic c stic
(Erythroblast)
Pronormobl Rubriblast Proerythrobl
ast ast  Nucleus
Basophilic Prorubicyte Basophilic o N:C ratio: 4:1
normoblast erythroblast o Chromatin: partially
Poychromat Rubricyte Polychroma
clumped, ‘wheel with broad
ic tic
spokes'
(polychrom
atophilic) o Nucleoli: ABSENT
erythroblast  Cytoplasm
Orthochtom Metarubicyt Orthochrom o Deeply basophilic
ic e ic o No granules
normoblast erythroblast
Polychroma Polychroma Polychroma Rubricyte/Polychromatic Normoblast
tic tic tic (Erythroblast)
(polychrom (polychrom (polychrom
 “dawn of hemoglobinization”
  Last stage capable of mitosis Erythrocyte
 Nucleus:
 No nucleus
o N:C ration 1:1
 Biconcave disc
o Chromatin: increasingly
 Salmon pink-staining cell with a
clumped
central pale area
o Nucleoli: ABSENT
 Area of central pallor of 1 to 3 um
 Cytoplasm (1/3 of the cell)
o Mixtures of the red-staining
 Circulate for about 120 days
of hemoglobin with the blue
of RNA in varying shades of
gray
o Pink coloration mixed with
basophilia: muddy, light
gray appearance.
Metarubricyte/Orthochromic Normoblast
(Erythroblast)
 Nucleated RBC
 Mitosis no longer possible
 Nucleus
o N:C ratio: LOW
o Chromatin: pyknotic (dense White Blood Cells
or compact)
o Nucleoli: ABSENT Granulocytic Series
 Cytoplasm
o Acidophilic (reddish pink)
cytoplasm
o Salmon-pink color
Polychromatic Erythrocyte
 Called reticulocytes when stained
with a vital stain
 No nucleus
 Polychromatophilic as a result of
the retention of RNA
 Synthesizing hemoglobin, for 2 to 3
days in the marrow and for 1 day Myeoblast
in the blood  Earliest morphologically
 Seen in Wright’s stain as large identifiable granulocytic precursor
bluish-red cells, polychromatophilic  Nucleus:
macrocytes
 o Shape: large oval
quadrangular nucleus
o N:C ratio: 4:1
o Chromatin: very
uniform, finely reticular
o Nucleoli: 1 – 5
 Cytoplasm:
o Small rim of basophilic
cytoplasm that lacks
granules, appears smooth and
velvety
Promyelocyte
 Production of azurophilic granules
(NON-SPECIFIC GRANULES)
 Nucleus
o Shape: oval and is often
eccentric. A paranuclear
halo or “hof”
o N:C ratio: 3:1
o Chromatin: more condensed
than in the blast
o Nucleoli: 1
 Cytoplasm
o Basophilic and is filled by
more and more azurophilic
granules
o Pale grayish blue
Myelocyte
 “dawn of neutrophilia”
 Last stage capable of cell division
 Nucleus:
o Shape: Oval indented
o N:C ratio: 2:1
o Chromatin: red purple with
slight granular, coarser,
clumped
 Cytoplasm:
to o Appearance of secondary or
specific cytoplasmic
granulation
fine, o Neutrophilic
 Fine and stain a
blue-pink color
o Eosinophilic
 Larger, oval shaped,
orange with glassy
texture
o Basophilic
 Dark blue-black
color and a dense
appearance
Metamyelocyte
 Synthesis of tertiary granules
 Color of the specific granulation
continues to become a major
distinguishing feature
 Nucleus
o Shape: indented or kidney
bean shape (peanut shape)
o N:C ratio: 1:1
o Chromatin: more condensed
or clumped
 Cytoplasm:
o Pale blue to pinkish tan with
moderate specific granules
Band/Stab
 Nucleus:
o Shape: Band shape like a
cigar band, C or S shaped,
unable to see filament
o N:C ratio: 1:1
o Chromatin: coarsely
clumped almost like leopard
spot coarseness
 Cytoplasm:
 o Brown-pink, with many fine
secondary granules
Mature/Segmented Neutrophil
 Nucleus:
o Shape: two to five lobes of
nucleus connected by thin
thread-like filaments
 Cytoplasm
o Pale lilac with blue shading
and many fine secondary
dust-like granules
o Life span: 6-10 hours
Basophil
 Can appear at the myelocytic
stages and move through the
maturation sequence
 Nucleus:
o Shape: coarse, clumped
bilobed
 Cytoplasm
o Many large SPECIFIC
purple-black granules seem
to obscure the large
Eosinophil cloverleaf form nucleus
o May decolorize during
 Can appear at the myelocytic
staining leaving pale areas
stages and move through the
within cell; granules much
maturation sequence
larger than neutrophilic
 Nucleus granules
o Shape: eccentric nucleus,
usually bilobed
 Cytoplasm
o Large, distinctive red-
orange SPECIFIC granules
with orange-pink
o Granules are uniformly
round, large, and
individualized; if stain is less
than adequate, observe
granules carefully for their
crystalloid nature White Blood Cells
Mononuclear Cells o Fine azure granules often
referred to as azure dust or a
ground-glass appearance
o Cytoplasmic and nuclear
vacuoles may also be present

Promonocyte
 Earliest recognizable cell in this
series
 Nucleus:
o Shape: oval or indented or
folded
o N:C ratio: 3:1
o Chromatin: fine uniform or
slightly streaked
o Nucleoli: 2-5
 Cytoplasm:
o Basophilic with a ground-
glass appearance and a
variable number of fine
azurophilic granules
Monocyte
 Nucleus:
o Shape: brainy convolutions
to lobulated and S shaped, Lymphoblast
horseshoe shaped
o N:C ratio: 1:1  Nucleus
o Chromatin: loose-weaved, o Shape: round
lacey, open, and thin o N:C ratio: 4:1
 Cytoplasm o Chromatin: smudgy
o Abundant gray-blue chromatin
o May show area of o Nucleoli: 1-2 (surrounded by
protrusion or blebbing dark rim of chromatin)
 Cytoplasm
 o Little, deep blue staining at o Cytoplasm is more
edge abundant with tendency for
azurophilic granules
Prolymphocyte
Mature B Cell (After Blast
 Nucleus Transformation)
o Shape: round
o N:C ratio: 3:1  Nucleus
o Chromatin: slightly o Shape: round or oval and
coarsened chromatin may be eccentrically placed
 Cytoplasm: o Chromatin: fine pattern
o Gray-blue, mostly blue at  Cytoplasm
edges o Nongranular
o Moderate in amount
o Mottled blue color
Small Lymphocyte
 Clumping od chromatin around the
nuclear membrane may help to
distinguish this from a nucleated Plasmacytoid Lympocytes
cell
 Nucleus:
 Resting cell
o Shape: round or oval and
 Nucleus:
may be eccentrically placed
o Shape: oval
o Chromatin: coarse and
o N:C ratio 4:1
irregularly spaced
o Chromatin: coarse lumpy
o Nucleoli: may be visible
chromatin with specific areas
 Cytoplasm
of clumping, a compact cell
o Distinctive dark blue with a
 Cytoplasm
lighter staining area, the hof,
o Usually just a thin border,
next to the nucleus
with few azurophilic, red
granules Plasma Cell
Large Lymphocyte  Nucleus:
o Shape: small and
 Nucleus
eccentrically placed
o Shape: oval
o Chromatin: condensed and
o N:C ratio 3:1
has a cartwheel
o Chromatin: Looser
configuration
chromatin pattern, more
o More than one nucleus may
transparent
be seen
 Cytoplasm
 Cytoplasm
o Dark blue
 o The hof area is usually visible Megakaryocytic Development
o Oval in outline and
 Endomitosis
abundant
o Nuclear division without
o Granules are absent, but
cytoplasmic division
vacuoles are common
 Burst-forming-unit megakaryocyte
(BFU-M)
o Most primitive progenitor
cell committed to
megakaryocyte lineage
Megakaryoblast
 Most immature cell
o Overlapping nuclear lobes
and a small amount of
basophilic cytoplasm
o High nuclear to cytoplasmic
ratio
o Two to six nucleoli
Other Forms of Plasma Cells
 Plasma membrane blebs
 Grape or Mott cells o Blunt projections from the
o Cytoplasm is completely margin that resemble platelets
filled with Russell bodies
Promegakaryocyte
 Flame cells
o The cytoplasm stains a  A large cell of 80 um with dense
bright-red color and alpha and lysosomal granules
contains increased  Nuclear lobularity first becomes
quantities of glycogen or apparent as an indentation
intracellular deposits of
amorphous matter Megakaryocyte
 Largest bone marrow cells
 NC ratio: 1:12
 Multilobular, not multinucleated
 Nucleus is more compact
 Basophilia has disappeared
 Granules are clustered into small
aggregates
Platelets
 No nucleus
 Light blue, with evenly dispersed,
fine red-purple granules
 Life span of 7 to 10 days
 Formed by proplatelet process
o Pseudopodial extensions of
megakaryocytes that
progressively branch and
thin out
o Develop from invaginated
surface membranes
(demarcation membranes)