HEMATOLOGY only in the vertebrae, ribs, sternum, skull bones, pelvis,
MODULE 3 & 4: HEMATOPOIESIS
AND LINEAGE SPECIFIC HEATOPOIESIS
Hematiopoiesis Defined
Hematopoiesis is the process of blood cell production,
differentiation, and development. The hematopoietic
system consists of the bone marrow, liver, spleen, lymph
nodes, and thymus.Before investigating the general
maturational characteristics of cells, knowledge of blood
cell development is useful.
Origin of Blood Cells
Hematopoietic stem cells (HSCs) are the foundation of
the adult hematopoietic system. It is now widely
accepted that the embryo produces the first adult
repopulating HSCs.
Types of Human Stem Cells
Functionally, three types of human stem cells exist:
1. Totipotential stem cells. These cells are present in the
first few hours after an ovum is fertilized. Totipotential
stem cells, the most versatile type of stem cell, can
develop into any human cell type, including
development from embryo into fetus.
2. Pluripotential stem cells. These cells are present
several days after fertilization. Pluripotent stem cells can
develop into any cell type, except they cannot develop
into a fetus.
3. Multipotential stem cells. These cells are derived
from pluripotent stem cells. They can be found in adults,
but they are limited to specific types of cells to form
tissues. For example, bone marrow stem cells can
produce all types of blood cells, bone cartilage, and
adipose (fat) cells.
Bone Marrow Sites and Functions
Bone marrow is found within the cavities of all bones
and maybe present in two forms: yellow marrow, which
is normally inactive and composed mostly of fat
(adipose) tissue, and red marrow, which is normally
active in the production of most types of leukocytes,
erythrocytes, and thrombocytes.
The bone marrow is one of the body’s largest organs. It
represents approximately 3.5% to 6% of total body
weight and averages around 1,500 g in adults, with the
hematopoietic marrow being organized around the bone
vasculature. The bone marrow consists of hematopoietic
cells (erythroid, myeloid, lymphoid, and
megakaryocyte), fat (adipose) tissue, osteoblasts and
osteoclasts, and stroma. Hematopoietic cell colonies are
compartmentalized in the cords. Following maturation in
the hematopoietic cords, hematopoietic cells cross the
walls of the sinuses, specialized vascular spaces, and
enter the circulating blood.
During the first few years of life, the marrow of all
bones is red and cellular. The red bone marrow is
initially found in both the appendicular and the axial
skeleton. In young persons but progressively becomes
confined to the axial skeleton and proximal ends of the
long bones in adults. By age 18, red marrow is found
and to some extent the proximal epiphyses of the femur
and humerus
Hematopoietic Development
 Hematopoiesis is a continuous, regulated
process of blood cell production that includes
cell renewal, proliferation, differentiation, and
maturation.
 During the fetal development, the restricted,
sequential distribution of cells initiates in the
yolk sac and then progresses in the aorta- gonad
mesonephros region (mesoblastic phase), then to
the fetal liver (hepatic phase), and finally resides
in the bone marrow (medullary phase).
Rodak’s Hematology Chapter 7 page 76
Mesoblastic phase
 Hematopoiesis is considered to begin around the
19th day of embryonic development after
fertilization. Early in embryonic development,
cells from the mesoderm migrate to the yolk sac.
 These primitive but transient yolk sac
erythroblasts are important in early
embryogenesis to produce hemoglobin (Gower-
1, Gower-2, and Portland) needed for delivery of
oxygen to rapidly developing embryonic tissues.
Rodak’s Hematology Chapter 7 page 76
Hepatic phase
 The hepatic phase of hematopoiesis begins at 5
to 7 gestational weeks and is characterized by
recognizable clusters of developing
erythroblasts, granulocytes, and monocytes
colonizing the fetal liver, thymus, spleen,
placenta, and ultimately the bone marrow space
in the medullary phase.
 Production of megakaryocytes also begins
during the hepatic phase. The spleen gradually
decreases granulocytic production and involves
itself solely in lymphopoiesis.
Rodak’s Hematology Chapter 7 page 77
Medullary phase
 Prior to the fifth month of fetal development,
hematopoiesis begins in the bone marrow cavity.
The transition is called medullary hematopoiesis
because it occurs in the medulla or inner part of
the bone.
Rodak’s Hematology Chapter 7 page 78
 Erythropoiesis – occurs in distinct anatomical
sites called erythropoietic islands, specialized
niches in which erythroid precursors proliferate,
differentiate, and enucleate.
 Granulopoiesis – Myeloid cells account for 23%
to 85% of the nucleated cells in normal bone
Page | 1
 marrow. Granulopoiesis can be recognized as a Length of Time in This Stage: This stage lasts
maturational unit. approximately 30 hours.
Orthochromic Normoblast (Metarubricyte)
 Lymphopoiesis – Unlike other cell lines,
lymphocytes and plasma cells are produced in Cytoplasm: The increase in the salmon-pink color of the
lymphoid follicles. Lymphocytes are randomly cytoplasm reflects nearly complete hemoglobin
dispersed throughout the cords. Lymphoid cells production.
typically account for 1% to 5% of the nucleated
cells in the normal bone marrow. Nucleus: The nucleus is completely condensed
N/C ratio: approx. 1:2
 Megarkayopoiesis – Megakaryopoiesis takes
place adjacent to the sinus endothelium. Length of Time in This Stage: This stage lasts
Megakaryocytes protrude through the vascular approximately 48 hours.
wall as small cytoplasmic processes to deliver Polychromatic (Polychromatophilic) Erythrocyte or
platelets into the sinusoidal blood. Reticulocyte
Megakaryocytes develop into platelets in
approximately 5 days. Cytoplasm: By the end of the polychromatic erythrocyte
stage, the cell is the same color as a mature RBC,
Turgeon - Clinical Hematology Theory & Procedures Chapter 4 page 79 salmon pink.
Three Erythroid Precursor Nomenclature Systems Nucleus: Beginning at the polychromatic erythrocyte
stage, there is no nucleus.
Length of Time in This Stage: The cell typically
remains a polychromatic erythrocyte for about 3 days
Erythrocyte
Cytoplasm: The mature circulating erythrocyte is a
Pronormoblast (Rubriblast) biconcave disc measuring 7 to 8 mm in diameter, with a
thickness of about 1.5 to 2.5 mm.
Cytoplasm: Dark blue
Nucleus: No nucleus is present in mature RBCs.
Nucleus: round to oval, containing one or two nucleoli.
Location and Length of Time in This Stage: Mature
The purple red chromatin is open and contains few, if
RBCs remain active in the circulation for approximately
any, fine clumps.
120 days
N/C ratio: 8:1 Rodak’s Hematology Chapter 8 page 98 - 103

Length of Time in This Stage: This stage lasts slightly Leukopoiesis

more than 24 hours.
Basophilic Normoblast (Prorubricyte)  Leukopoiesis is stimulated by interleukins and
colonystimulating factors (CSFs)
Cytoplasm: When stained, the cytoplasm may be a
 Macrophages and T cells are the most important
deeper, richer blue than in the pronormoblast hence the
sources of cytokines.
name basophilic for this stage
Granulopoiesis
Nucleus: The chromatin begins to condense, revealing
clumps along the periphery of the nuclear membrane and
a few in the interior
N/C ratio: 6:1
Length of Time in This Stage: This stage lasts slightly
more than 24 hours.
 Granulocytes are a group of leukocytes whose
Polychromatic (Polychromatophilic) Normoblast
cytoplasm is filled with granules with differing
(Rubricyte)
staining characteristics and whose nuclei are
Cytoplasm: This is the first stage in which the pink segmented or lobulated. Individually, they
color associated with stained hemoglobin can be seen. include eosinophils, with granules containing
basic proteins that stain with acid stains such as
Nucleus: The chromatin pattern varies during this stage eosin; basophils, with granules that are acidic
of development, showing some openness early in the and stain with basic stains such as methylene
stage but becoming condensed by the end. The blue; and neutrophils, with granules that react
condensation of chromatin reduces the diameter of the with both acid and basic stains, which gives
nucleus considerably. them a pink to lavender color.
N/C ratio: from 4:1 to about 1:1 at the end of this stage

Page | 2
  Neutrophils – Also known as
polymorphonuclear cells or PMNs, neutrophils
are present in the peripheral blood in two forms
according to whether the nucleus is segmented
or still in a band shape. Segmented neutrophils
make up the vast majority of circulating
leukocytes.
 Eosinophils - Eosinophils make up 1% to 3% of
nucleated cells in the bone marrow. Of these,
slightly more than a third are mature, a quarter
are eosinophilic metamyelocytes, and the
remainder are eosinophilic promyelocytes or
eosinophilic myelocytes. Eosinophils account
for 1% to 3% of peripheral blood leukocytes,
with an absolute number of up to 0.4 3 10/L in
the peripheral blood.
 Basophils - Basophils and mast cells are two
cells with morphologic and functional
similarities; however, basophils are true
leukocytes because they mature in the bone
marrow and circulate in the blood as mature
cells with granules, whereas mast cell precursors
leave the bone marrow and use the blood as a
transit system to gain access to the tissues where
they mature. Basophils are discussed first.
Basophils are the least numerous of the WBCs,
making up between 0% and 2% of circulating
leukocytes and less than 1% of nucleated cells in
the bone marrow.
Rodak’s Hematology Chapter 12 page 149 to 158
Normal Maturation Characteristics of Neutrophils,
Eosinophils, and Basophils.
Myeloblast - the earliest morphologically identifiable
granulocytic precursor is the myeloblast. This cell has an
average overall diameter of 10 to 18 mm. The nuclear
chromatin is finely reticular, with one to five light-
staining nucleoli. The cytoplasm appears as a small rim
of basophilic cytoplasm that lacks granules.
Promyelocyte - represents the second maturational stage
seen in granulocytes. The outstanding feature of this cell
is the presence of prominent granulation that may
actually obscure the other morphological features of the
cell. These granules are primarily azurophilic granules
and are rich in the enzyme myeloperoxidase and
chloroacetate esterase.
Myelocyte – Is the third maturational stage. This cell is
characterized by the recognizable appearance of
secondary or specific cytoplasmic granulation. These
round or oval-shaped granules are orange and have a
glassy or semi opaque texture. Basophilic granules have
a dark blue-black color and a dense appearance. The
myelocyte has an average diameter of 12 to 18 um.
Metamyelocyte – Is the fourth maturational stage. Its
most characteristic feature is that the nucleus begins to
assume an indented or kidney bean shape, which will
continue to elongate as the cell matures through this
phase. The chromatin continues to become more
condensed or clumped. The color of the specific
granulation continues to become a major distinguishing
feature.
Mature Forms
Two stages of granulocytes are observed in the
circulating blood: the band form and the segmented
form. The band form has a typical elongated nucleus.
Turgeon - Clinical Hematology Theory & Procedures Chapter 14 page 237 to page 239
Lymphopoiesis
 Lymphocytes are derived from committed stem
cells that originate from pluripotent stem cell.
 Ealy lymphoid cells further differentiates into B
& T lymphocytes.
 T-cell maturation happens in the Thymus while
B-cell maturation happens in the bone marrow.
Primary lymphoid
organs
Secondary lymphoid
organs
Lymphocytes
 Bone marrow  Lymph nodes  B & T
 Thymus  Spleen lymhpocytes
 Lymphoid  NK cells
tissues
Lymphoblast - is the first morphologically identifi able
cell of the lymphocytic maturational series in the bone
marrow. The overall size ranges from 15 to 20 mm, with
a nuclear-cytoplasmic (N:C) ratio of 4:1.
Prolymphocyte - The second stage in the maturational
development of the lymphocyte is the prolymphocyte.
This cell may be seen in the bone marrow, thymus, and
secondary lymphoid tissues. The overall size is usually
about the same (15 to 18 mm) as the lymphoblast. The
N:C ratio ranges from 4:1 to 3:1.
Mature Lymphocyte - range in size from large (17 to 20
mm) in younger cells to small (6 to 9 mm) in older cells.
Page | 3
The N:C ratio ranges from 2:1 in younger cells to 4:1 to Types of Leukocytes
3:1 in older cells. Granulocytes Agranulocytes
 Neutrophils (40-70%)  Monocytes(4-8%)
Turgeon - Clinical Hematology Theory & Procedures Chapter 16 page 269
 Eosinophils(1-4%)  Lymphocytes(20-
 Basophils(<1%) 45%)

Movement of cells

Chemotaxis - Cells are guided to the site of injury by the

concentration gradient of chemotactic substances.
Diapedesis - The marginating pool of WBCs, adhering
to the endothelial lining of capillaries, migrates through
the vessel wall to the intestinal tissues.

Turgeon - Clinical Hematology Theory & Procedures Chapter 14 page 246

Thrombopoiesis

Megakaryoblast – The cell is large, irregularly shaped

with a single or several round or oval nuclei and with a
Monopoiesis
blue, non-granular cytoplasm. Nucleoli are usually
present.
Promegakaryocyte – This cell differs from the
megakaryoblast in that there are bluish granules in the
cytoplasm.
Megakaryocyte – The cell is very large with relatively
large amounts of cytoplasm, and multiple nuclei. The
cytoplasm contains numerous small, uniformly
distributed granules that are reddish-blue in color.

Monoblast
Size(um): 12-20
Stages in Thrombocyte development
N/C ratio: 4:1
Chromatin: Fine
Three stages of Maturation of megakaryocytes
Shape: Oval, folded
Color: Blue 1. Basophilic stage – Megakaryocyte is small, has
diploid nucleus and abundant basophilic
Promonocyte
cytoplasm.
Size(um): 12-20 2. Granular stage – the nucleus is more polypoid,
N/C ratio: 3:1-2:1 cytoplasm is more eosinophilic and granular.
Chromatin: Lace like 3. Mature stage – megakaryocyte is very large,
Shape: Elongated, folded with approx. 16-32 nuclei, abundance of
Color: Blue-gray granular cytoplasm.

Mature Monocyte Normal Cellular Maturation

Size(um): 12-18
N/C ratio: 2:1-1:1 1. Cytoplasmic Maturation
Chromatin: Lace like  Immature cell: intensely basophilic
Shape: Horseshoe shaped, folded  Granules may appear as cell matures
Color: Blue-gray 2. Nuclear Maturation
 Immature nucleus is round or oval and
Turgeon - Clinical Hematology Theory & Procedures Chapter 14 page 242
large in proportion to rest of the cell.
 As cell matures, it decreases in size and
takes on varying shapes.
 Nuclear chromatin changes from
delicate and fine to coarse and clumped.
Page | 4
  Staining properties change frome
reddish purple to bluish purple.
 Nucleoli gradually disappear.
3. Cell Size
 As cell matures, it becomes smaller in
size.
Megakaryopoiesis
Megakaryopoiesis takes place adjacent to the sinus
endothelium. Megakaryocytes protrude through the
vascular wall as small cytoplasmic processes to deliver
platelets into the sinusoidal blood. Megakaryocytes
develop into platelets in approximately 5 days.
Other Cells Found in Bone Marrow
Marrow Stromal Cells
The meshwork of stromal cells is composed of reticulum
cells, histiocytes, adipose cells, and endothelial cells.
This structure is where the hematopoietic cells are
suspended in a delicate semi-fluid state. Stromal cells
produce an extracellular matrix composed of collagens
and proteins (e.g., glycoproteins and proteoglycans). The
extracellular matrix is critical for the maintenance of
normal renewal and differentiation of bone marrow cells.
Mast Cells
Tissue mast cells, a connective tissue cell of
mesenchymal origin, are normally observed in bone
marrow. The abundant blue-purple granules that usually
obscure the round or oval reticular nucleus contain
heparin, histamine, serotonin, and proteolytic enzymes.
Increased numbers of mast cells can be seen in a variety
of abnormal conditions (e.g., chronic
lymphoproliferative disorders or chronic infections).
Macrophages
Macrophages, also called reticulum cells or histiocytes,
appear as large cells in the bone marrow. The
appearance of the cytoplasm will vary, depending on
what the cell has ingested (e.g., siderophages are
macrophages containing iron-rich hemosiderin and
ferritin). Gaucher cells are macrophages filled with
uncatabolized glucocerebrosides.
Bone Cells
Osteoblasts are bone matrix–synthesizing cells that
resemble plasma cells and are usually observed in
groups. Although these cells are only occasionally seen
in normal adult bone marrow aspirates, an increased
number of cells is characteristically seen in aspirates
from children and from patients who have metabolic
disease. Osteoclasts resemble megakaryocytes. These
are bone-remodeling cells.
Interlukins
The interleukins were first described as signals for
communication between (inter—between) white blood
cells (leuk—from leukocytes). Currently, it is well-
known that these molecules are produced and used as
signaling molecules in many cells of the body, in
addition to immune cells.
Interleukins are basically the method of immune cross-
talk and communication. Interleukins are the primary
messengers and directors of the immune system. There
are currently 35 well-known interleukins; however, there
are many more to be found and characterized. Acting
network of inflammatory stimuli and cytokines suggests
that these growth factors may have a limited role in
hematopoietic homeostasis but a major role in host
responses to infection or antigenic challenge. They can
cause cellular proliferation, cell activation,
inflammation, physiology changes such as fever and
pain, and allergies as with histamine release and growth.
Page | 5
Page | 6