m02 Hematopoiesis 1 2

MODULE 2
HEMATOPOIESIS
UNIT 1: INTRODUCTION TO HEMATOPOIESIS
At the end of the module, students should be able to:
1. Discuss the maturation sequence of blood cells from embryo to fetus to adult.
2. Differentiate immature from mature blood cells.
3. Describe the microscopic presentation of the different blood cells.
4. Discuss the roles of hematopoietic growth factors in blood cell production.
Hematopoiesis (Fig. 2-1) is the process of cellular

formation, proliferation, differentiation and maturation
of blood cells.
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Figure 2-1. Overview of Cellular Maturation
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PHASES OF HEMATOPOIESIS (Fig. 2-3)
Hematopoiesis is generally categorized as primitive or definitive. Primitive
hematopoiesis occurs in the embryo during the first two weeks and lasts up to the
eighth week of gestation. It is the time when the blood cells produced are mostly
primitive erythrocytes. Definitive hematopoiesis occurs on the eighth week until
adulthood. In this stage, different blood cells are produced which can be distinguished
morphologically and functionally thus the term definitive.
1. Mesoblastic Phase (Fig. 2-2)

 “Yolk sac phase”
 Begins around 19th day of embryologic development
 PRIMARY SITE OF HEMATOPOIESIS: Blood islands of the YOLK SAC
(Hematopoiesis occurs intravascularly)
i. Cells of the yolk sac
1. Mesodermal cells: Develop to primitive erythroblasts
2. Angioblasts: Forms the future blood vessels
 BLOOD CELL/S FORMED:
i. Erythroblasts (1st month of embryonic development)
 Embryogenic hemoglobins are formed:
i. Gower I (2 zeta chains & 2 epsilon chains)
ii. Portland (2 zeta chains & 2 gamma chains)
iii. Gower II (2 alpha chains & 2 epsilon chains)
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Figure 2-2. Embryonic hematopoiesis
2. Hepatic Phase
 Begins at around 5-7 gestational weeks
 PRIMARY SITE/S OF HEMATOPOIESIS: FETAL LIVER
1. Liver
 Becomes the primary site of hematopoiesis during the 3rd month of
fetal development
 Retains minimal activity up to 1-2 weeks after birth
2. Thymus
 First fully developed organ in the fetus
 Becomes the major site of T cell production
3. Spleen & Kidneys
 Production of B lymphocytes
 Spleen gradually decreases granulocytic production and involves itself
solely in lymphopoiesis
4. Lymph nodes
 BLOOD CELL/S FORMED:
 Erythrocytes still in production
 Granulocytes & Megakaryocytes (3rd month of gestation)
 Lymphocytes (4th month of gestation)
 Monocytes (5th month of gestation)
 FETAL HEMOGLOBIN (4th month of gestation) is the PREDOMINANT
HEMOGLOBIN but detectable levels of adult hemoglobin may be present
i. HbF: 2 alpha & 2 gamma chains
Figure 2-2. Phases of Hematopoiesis
3. Myeloid Phase
 Begins prior to the 5th month of development
 PRIMARY SITE/S OF HEMATOPOIESIS: BONE MARROW (end of 6th month)
i. “Medullary hematopoiesis”: Hematopoiesis occurs inside the medulla of
the bone (where the bone marrow is located) (Fig. 2-4).
In the space provided, illustrate the cross-section of a long bone and label the parts. Take
note of the location of the medullary cavity. (See Keohane, Smith, and Walenga, 2016
for reference.)
ii. At birth, bone marrow becomes the ONLY SITE FOR PRODUCTION of
erythrocytes, granulocytes, monocytes, platelets, and B lymphocytes
 Myeloid-to-erythroid ratio gradually approaches 3:1 (adult levels)
 Measurable levels of Hemoglobins F and A
i. HbA : 2 alpha chains & 2 beta chains
ii. HbA2: 2 alpha chains & 2 delta chains
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Figure 2-4. Sites of Hematopoiesis
Adult Hematopoietic Tissues

- Adult hematopoietic tissues can be classified according to their roles in lymphocyte
development.
A. Primary lymphoid tissues

The primary lymphoid tissues function for the production and maturation of T
and B lymphocytes. These include the bone marrow and the thymus.
1. BONE MARROW (BM)

2 types:
 Yellow marrow: Consists primarily of adipocytes
 Red marrow: Hematopoietically active
Figure 2-5. Myeloid Hematopoiesis
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Figure 2-5 shows that during infancy and early childhood; all bones in the body contain
the red marrow. By age 5 to 7, retrogression occurs. This is the process of replacing the
haematopoietically active red marrow with yellow marrow. The yellow marrow is consisting
of adipocytes that is capable of reverting back to active marrow in cases of increased
demand for blood cell production in the body.
ACTIVITY 1 (MYELOID HEMATOPOIESIS): From the illustration of the skeletal system

provided, compare and contrast the different bones of an adult to that of an
infant in terms of their function of blood cell production: (5 points)
Structure of the BM (Fig. 2-6):
A. Vascular region: Vascular sinuses
 Vascular sinuses (specialized blood vessels)
B. Hematopoietic cords
These are extravascular cords that are composed of hematopoietic cells
and macrophages. They are located in spaces between the vascular
sinuses and are supported by trabeculae of spongy bone. It is noted that
hematopoietic cells develop in specific niches within the cords.
C. Trilaminar sinus wall

The trilaminar sinus wall separates the extravascular cords from the vascular
sinuses (Fig. 2-7).
 Components (From the extravascular cords to the vascular sinus)
a. (Reticular) Adventitial cells
 Incomplete layer of cells on the abluminal surface of the vascular sinus
(facing the extravascular cords)
b. Basement membrane
c. Endothelial cells
 Form a single, continuous layer along the luminal (inner) surface of
vascular sinuses
Niches/ Hematopoietic
Microenvironment (within the
cord) play a very important role
in nurturing and protecting
hematopoietic stem cells
Erythroblast:
 Develop in small
clusters; More mature
forms located in outer
surfaces of the vascular
sinuses
 Found surrounding
iron- laden
macrophages
Megakaryocytes: Adjacent to
the walls of vascular sinuses
Immature myeloid cells (up to
metamyelocyte stage): Deep
within the cords (as the
Figure 2-6. Cross-section of bone with active marrow mature, they move closer to
the vascular sinuses)
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Figure 2-7. Histology of the red bone marrow
ENTRY OF MATURE BLOOD CELLS FROM THE BONE MARROW TO PERIPHERAL CIRCULATION
Mature blood cell  Adventitial cell layer (contracts)  Basement membrane  Endothelial
cell layer  Receptor-mediated process  Mature cells bind to the surface of
endothelial cells  Cells pass through pores in the endothelial cytoplasm  Vascular sinus
 Peripheral circulation
Bone Marrow Specimens
Collection
1. Trephine/ Core biopsy
 Utilizes Trephine biopsy needle (Jamshidi needle)
2. Aspiration
 Aspiration needle (University of Illinois sternal needle)
Collection is oftentimes carried out to observe the patient’s myeloid-to-erythroid ratio

Myeloid-to-Erythroid ratio:
Normal: 2:1 to 4:1 (Average of 3:1)
Infection: 6:1
Leukemia: 25:1
Normal marrow cells
a. All developing hematopoietic cells
b. Macrophages
c. Mast cells
d. Osteoblasts
 Waterbug or comet appearance
 Confused with plasma cells
e. Osteoclasts
 Misidentified as megakaryocytes
2. THYMUS
Figure 2-8. The anatomy (left) and histology (right) of the thymus.
The thymus is a small, flat bilobed organ (Fig. 2-8, left) found in the thorax that has
an average 30g weight at birth, 35 g weight at puberty and gradually atrophies.
It is where T-cell maturation happens.
- Parts (Fig. 2-8, right):

 Capsule
The capsule separates the two lobes of the thymus. It has
extensions known as Trabeculae that penetrate the thymus and divides the
two lobes into lobules
 Lobules
 Cortex
It is the outer part of the thymic lobule and consists of a
large amount of pre-T cells and scattered dendritic cells, epithelial
cells, and macrophages.
Pre-T cells are immature T cells that migrate from the red
marrow to the thymic cortex and proliferate and start to mature at the
cortex.
Dendritic cells are derived from the monocytes. They exhibit
long dendrite-like projections and assist the maturation process of the
pre-T cells.
Epithelial cells are specialized to carry out the positive selection
process of pre-T cells. They have long processes that serves as a
framework for the T cells and produce thymic hormones that are
thought to aid in the maturation of T cells
The thymic macrophages help clear out the debris of dead
and dying cells.
 Medulla
The medulla is the inner part of the thymic lobule. It consists of
more mature T cells, dendritic cells, and epithelial cells, and
macrophages.
In the medulla are* Thymic (Hassall’s) corpuscles which are
clusters of epithelial cells that become arranged into concentric layers
of flat cells that degenerate & become filled with keratohyalin granules
and keratin.
B. Secondary lymphoid tissues

These tissues are where lymphoid cells become competent (where lymphoid
cells respond to foreign antigens). These consist of the spleen, lymph nodes,
and lymphatic nodules.
1. Lymph Nodes
Lymph nodes (Fig. 2-9) are bean-shaped structures found along lymphatic
vessels which are specialized to filter lymph flowing through the lymphatic
vessels.
Figure 2-9. Structure of the lymph node

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Parts:
 Cortex
The cortex contains the primary and secondary follicles. The primary
follicles contain mature B cells, follicular dendritic cells, and macrophages. They
are located are B cells that are not yet introduced and stimulated by any antigen
thus become the site of antigen recognition of the mature B cells. The secondary
follicles (B-cell area) arises from the primary follicles after its B cells have
recognized a specific antigen. It has a central portion known as germinal center
which is the site of blast transformation of B cells and also of plasma cell and
memory B cell formation.
 Paracortex (T-cell area)

This is the region between cortex and the medulla. It contains
structures known as high endothelial venules which are specialized
venules in the paracortex where numerous lymphocytes enter from the
bloodstream. They are composed mainly of T cells and antigen presenting
cells known as the interdigitating cells.
 Medulla
The medulla is less densely populated and contains T cells, B cells,
macrophages, and numerous plasma cells.
2. Spleen
The spleen is the largest secondary lymphoid organs that functions as a large
discriminating filter. It removes damaged cells and foreign antigens from the blood.
Figure 2-10. Structure of the Spleen
- Parts of the Spleen (Fig. 2-11)
 Stroma
The stoma consists of capsule, trabeculae, reticular fibers and
fibroblasts.
 Parenchyma
The parenchyma is the functional part of the spleen. It consists of two
different kinds of tissue. The white pulp is approximately 20% of the
weight of the spleen and consists of lymphoid tissues. The structures of
the white pulp are the following:
i. Periarteriolar lymphoid sheath (PALS)
- Contains mainly T cells
ii. Primary follicles
- Contains B cells that are not yet introduced and
stimulated by the antigen
iii. Marginal Zones
- Surrounds the primary follicles
- Contains dendritic cell that traps antigens
iv. Germinal Centers in secondary follicles
- Site of blast transformation of B cells
- Site of formation of plasma cells and B memory
cells The red pulp makes up more than one half of the volume of
the spleen. It has the following parts:
i. Venous sinuses
- Blood-filled structures
ii. Splenic (Billroth’s) Cords
- Cords of splenic tissue
- Consists mainly of:
 Red blood cells
 Macrophages
 Lymphocytes
 Plasma cells
 Granulocytes
3. Lymphatic nodules
The lymphatic nodules are egg-shaped masses of lymphatic tissue that resembles
lymph nodes but do NOT have capsules. They may be present in:
 Small, solitary form
i. Mucosa associated lymphoid tissue
- Scattered in the lamina propria of the mucosa of the gastrointestinal,
urinary and the reproductive tracts
ii. Cutaneous associated lymphoid tissue
- Intraepidermal lymphocytes (mostly T cells)
 Multiple, aggregated form
i. Tonsils
ii. Peyer’s patches
- Aggregated lymphatic follicles in the ileum of the small intestine
STEM CELL THEORY
STEM CELLS
Stem cells are undifferentiated/ slightly differentiated cells that may either self-
renew or give rise to cells of different lineages
- Types:
a. Totipotential stem cell that can differentiate into all possible cells of the
organism, including the extra-embryonic membranes
b. Pluripotential stem cell that can give rise to all of the cells of the embryo, and
therefore of a whole animal, but are no longer capable of giving rise to extra-
embryonic structures
c. Multipotential stem cell that can give rise to multiple lineages but has lost the
ability to give rise to all body cells
HEMATOPOIETIC STEM CELL THEORY
1. Monophyletic theory
The monophyletic theory states that blood cells are derived from a single
progenitor cell (Pluripotent hematopoietic stem cell. It is the most widely
accepted theory
2. Polyphyletic theory
It states that each blood cell lineages are derived from own unique stem cell.
STEM CELL DIVISION

Phase Activity
Interphase
Period between cell divisions; chromosomes not visible under the light microscope
G0 phase Limbo phase; Cells that are not dividing and possibly never to divide
again
G1 phase (8-10 Metabolically active cell duplicates most of its organelles and
hours) cytosolic components
Replication of chromosome begins
S phase (8 hours) Replication of DNA and chromosomes

G2 phase (4-6 Cell growth, enzyme and protein synthesis continue
hours) Replication of centrosome complete
Mitotic Phase
Parent cell produces identical cells with identical chromosomes; chromosomes visible
under the light microscope
Mitosis
 Nuclear division
 Distribution of two sets of chromosomes into separate
nuclei Prophase Chromatin fibers condense into paired
chromatids
Nucleolus and nuclear envelope disappear
Each centrosome moves to an opposite pole of the cell
Metaphase Centromeres of chromatid pairs line up at the metaphase plate
Anaphase Centromeres split
Identical sets of chromosomes move to opposite poles of
cell Telophase Nuclear envelopes and nucleoli reappear
Chromosome resume chromatin form
Mitotic spindle disappears
Cytokinesis
 Cytoplasmic division
 Usually begins in late anaphase with the formation of a cleavage furrow & is
completed after the telophase
After completing the cell cycle, the stem cells have the following possible fates:
1. Apoptosis
 Programmed cell death
2. Self-renewal
 Returning of daughter cell to stem cell pool
3. Differentiation
 Stem cells differentiate to acquire new morphologic features and give rise to
more mature forms
Types of Division
1. Symmetric division
 Both daughter cells follow the path of differentiation
2. Asymmetric division
 One daughter cell returns to stem cell pool while the other differentiates
Models of Stem Cell Fate

1. Stochastic model
 Random commitment of stem cells to either self-renew or differentiate
2. Instructive model
 Signals from the hematopoietic inductive microenvironment determine the
fate of the hematopoietic stem cell
3. Current model
 Incorporate both stochastic and instructive model
 Initial decision follows stochastic model while lineage differentiation follows
instructive model
Development
1. Synchronous development
 Cytoplasm and nucleus mature at the same rate
2. Asynchronous development
 Cytoplasm or nucleus mature first before the other
 Can lead to abnormality in shape and size
Cell Maturation
1. Blast cells do not have granules
2. Blast cells contain a large nucleus (3/3 to 7/8 of cell area) and a small amount
of cytoplasm
3. As cells mature, the cytoplasm becomes less basophilic (Exception: Plasma cell)
4. As cells mature, the chromatin of the nucleus becomes heavier, and the darker the
nucleus stains the heavier the chromatin is
5. As the cells mature, they become smaller (Exception: Megakaryocyte)
6. Nucleoli tend to disappear in mature cells
7. As cells mature, specific granules become less prominent and smaller
8. There are four different types of granules: neutrophilic, basophilic, eosinophilic, and
azurophilic (primary).
CYTOKINES AND GROWTH FACTORS
Cytokines
Cytokines are group of specific glycoproteins secreted by cells. In hematopoiesis,
they regulate the proliferation, differentiation, and maturation of hematopoietic
precursor cells. These include interleukins, lymphokines, monokines, interferons,
chemokines, and colony-stimulating factors (CSF).
Interleukins
 Interleukins are cytokines with multiple actions and are numbered by
scientists in the order in which they were identified.
 Characteristics:
a. They are proteins that exhibit multiple biologic activities, such as regulation
of autoimmune and inflammatory reactions and hematopoiesis
b. They have synergistic interactions with other cytokines.
c. They are part of interacting systems with amplification potential.
d. They are effective at very low concentrations
Colony-stimulating Factors (CSF)

CSFs have high specificity for their target cells and are active at low
concentrations.
The names of the individual factors indicate the predominant cell lines that
respond to their presence.
 Examples:
 G-CSF (Granulocyte Colony-stimulating Factor)
 Stimulates the proliferation of the granulocytic cell line
 GM-CSF (Granulocyte-Macrophage Colony-stimulating Factor)
 Stimulates the proliferation of the granulocytic-monocytic cell
line
 Also works synergistically with Interleukin-3 (IL-3) to
enhance megakaryocyte colony formation
Growth factors can be classified according to the part of the development

process that they influence.
a. Early-acting growth factors
 Multilineage in action
 Notable examples:
i. KIT ligand
 Also known as stem cell factor (SCF)
 An early-acting growth factor which attaches to the receptor
transmembrane KIT
 Binding of KIT ligand to the KIT receptor triggers the cell to
proliferate; Activation of the KIT receptor which is essential in
the early stages of hematopoiesis.
ii. FLT3 ligand

 Works synergistically with IL-3, GM-CSF, and other cytokines to
promote early hematopoietic stem cell proliferation and
differentiation.
 Regulates blood cell production by controlling the production,
differentiation, and function of granulocyte and
macrophages
iii. GM-CSF
 Induces expression of specific genes that stimulate
hematopoietic stem cell differentiation to the common myeloid
progenitor.
iv. Interleukin-3 (IL-3)
 Aka Multi-CSF
Figure 2-11. DERIVATION OF BLOOD CELLS FROM HEMATOPOIETIC STEM CELLS
(Source: Henry’s Clinical Diagnosis and Management by Laboratory Methods, 22nd Edition)
UNIT 2: ERYTHROPOIESIS
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Figure 2-12. Significant changes in blood cell maturation
EXPLAIN
ACTIVITY 2 (CELL MATURATION): According to the general rules (CAP Guidelines) of

cell maturation and the illustration (Fig. 2-12) provided above, explain the trends
(important events per stage) of erythrocyte maturation in terms of the following:
(5 points)
4. NUCLEAR CHANGES
5. CYTOPLASMIC CHANGES
6. CELL SIZE
Erythropoietin (EPO)
EPO is the glycoprotein hormone produced by the kidneys (renal peritubular
interstitial cells). Its main effect is to place more erythrocytes into circulation at a
faster rate by:
o Early release of reticulocytes
o Prevent apoptotic cell death
o Reduces maturation time inside bone marrow
Maturation sequence
I. Erythroid Progenitors
a. Pluripotential hematopoietic stem cell
b. CFU-GEMM/ CFU-S
c. CFU-MegE
d. *BFU-E (Particularly produced under increased demand for RBCs/ pathologic
erythropoiesis)
e. CFU-E
II. Erythroid Precursors
a. Pronormoblast
b. Basophilic normoblast
c. Polychromatic (polychromatophilic) normoblast
d. Orthochromic normoblast
e. Reticulocyte/ Polychromatic (polychromatophilic) erythrocyte
f. Erythrocyte
**Three Erythroid Precursor Nomenclature Systems

NORMOBLASTIC RUBRIBLASTIC ERYTHROBLASTIC
Pronormoblast Rubriblast Proerythroblast
Basophilic normoblast Prorubricyre Basophilic erythroblast
Polychromatic Rubricyte Polychromatic
(polychromatophilic) (polychromatophilic)
normoblast erythroblast
Orthochromic normoblast Metarubricyte Orthochromic erythroblast
Reticulocyte/ Reticulocyte/ Reticulocyte/
Polychromatic Polychromatic Polychromatic
(polychromatophilic) (polychromatophilic) (polychromatophilic)
erythrocyte erythrocyte erythrocyte
Erythrocyte Erythrocyte Erythrocyte
Approximately 18 TO 21 DAYS are required to produce a mature RBC from the BFU-E
o 1 week for BFU-E to mature to CFU-E
o Another 1 week for CFU-E to mature to pronormoblast
o Another 6-7 days for the precursors to mature enough to enter the circulation
o 8 to 32 mature RBCs usually result from a single pronormoblast
Erythroid Precursors - Keohane, Smith and Walenga, 2016
Figure 2-13. Red blood cell maturation sequence
Image from: https://www.google.com/search?q=eryhtropoiesis&tbm=isch&ved=2ahUKEwi6prCNlZXrAhUFHKYKHVFmC6sQ2-cCegQIABAA&oq=eryhtropoiesis&gs_lcp=CgNpbWcQAzIGCAAQChAYOgQIABBDOgUIABCxAzoCCABQu-

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UNIT 3: ERYTHROCYTE KINETICS
THE RBC MEMBRANE

Crucial to the red cell function is the structure of the red cell membrane (Fig. 2-14). It is
made up of proteins (52%), lipids (40%) and carbohydrates (8%). The membrane
needs to be flexible, deformable and semi-permeable so that the red cell will be able to
travel through the largest blood vessels to the smallest capillaries in order to deliver
oxygen to the farthest areas of the body. The red cell membrane (Fig. 2-15) is exhibited by
a fluid mosaic model – a continually moving sea of fluid lipids that contains a mosaic of
different proteins. Some proteins float freely like iceberg in the lipid seam whereas
others are anchored at specific parts.
Figure 2-14. Structure of the red cell membrane

Image from: https://www.google.com/search?q=structure+of+the+red+cell+membrane&tbm=isch&ved=2ahUKEwjhwuGhlpXrAhUQHKYKHbc1Cg8Q2-
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AQE&sclient=img&ei=bp4zX-HTGZC4mAW366h4&bih=657&biw=1349&rlz=1C1RLNS_enPH904PH904&hl=en&hl=en#imgrc=TPRUKlQG5b6oZM
Figure 2-15. Parts of the Red Cell Membrane
The red cell membrane can be divided into two parts – the lipid bilayer and the membrane
proteins. The lipid bilayer consists of two back-to-back layers made up of three types of
lipid molecules:
i. Phospholipids (75%)
Phospholipids are amphipathic lipids – they have both polar and
nonpolar parts. The polar part is the phosphate-containing “head”
and the nonpolar part has two long fatty acid “tails”. They can be
asymmetrically divided:
 Outer layer: Phosphatidylcholine & Sphingomyelin
 Inner layer: Phosphatidylserine
& Phosphatidylethanolamine
ii. Cholesterol (20%)
It is a steroid with an attached hydroxyl group, weakly amphipathic
and are interspersed among the other lipids in both layers of the
membrane. The polar part contains the hydroxyl group and the non-polar
part contains the steroid rings and hydrocarbon tail.
iii. Glycolipids (5%)
Glycolipids are lipids with attached carbohydrate groups that
appear only in the membrane layer that faces the extracellular
fluid. It is one of the reasons the two sides of the bilayer are
asymmetrical. The polar part contains the carbohydrate “Head”
and the nonpolar part contains the fatty Acid “Tail”.
The membrane proteins are of two (2) types. The peripheral proteins which are not
firmly attached in the membrane but rather attached to the polar heads of
membrane lipids and the integral proteins which are firmly attached to the bilayer
membrane and extend into or through the lipid bilayer among the fatty acids.
The glycocalyx is an extensive sugary coat made up of the carbohydrate portions of

the glycolipids and glycoproteins. It acts like a molecular signature, enables cells to adhere
to one another in some tissues and protects the cells from being digested by enzymes in
the extracellular fluid.
RBC METABOLISM
1. Embden-Meyerhof Pathway (EMP)

The EMP (Fig. 2-16) is the major source of red cell energy. It is the pathway
responsible for 90% of glycolysis carried out by RBCs. In the process of
glycolysis/ glucose catabolism, glucose is converted to pyruvate and the
resulting pyruvate can be metabolized either via:
 Aerobic pathway: Tricarboxylic acid cycle
- NOT used by RBC metabolism (absence of mitochondria)
- Pyruvate is converted to acetyl-coenzyme A (acetyl-CoA)
- For each mole of glucose, a total of 38 ATP molecules are
produced
However, 2 ATP molecules are needed to initiate respiration so
there is a net of 36 ATPs.
 ANAEROBIC PATHWAY/ ANAEROBIC GLYCOLYSIS

- Pyruvate is converted to lactic acid (reaction is catalysed by
LDH)
- For each mole of glucose, a total of 4 ATP molecules are
produced. However, for anaerobic glycolysis to occur, 2 moles
of ATP must be consumed resulting in a net gain of 2 ATP
moles
2. Hexose Monophosphate Pathway/ Pentose Phosphate Shunt

The PPP contributes to10% of glycolysis. It provides adequate stores of
NADPH needed to maintain GLUTATHIONE IN ITS REDUCED FORM to prevent
denaturation of hemoglobin. The main enzyme: Glucose-6-phosphate
dehydrogenase (G-6PD)
**G-6-PD deficiency often yields in the presence of Heinz bodies
3. Methemoglobin Reductase Pathway

- Maintains hemoglobin iron in Fe2+ (Ferrous state) to be functional
4. Rapoport- Leubering Pathway

- Responsible for generation of 2,3-DPG which regulates hemoglobin affinity for
O2
RBC DESTRUCTION
Due to natural catabolism, red blood cells will eventually experience deterioration
of their enzymes. As nonnucleated cells, the mature RBCs are unable to generate or
replenish enzymes including glycolytic enzymes that lead to senescence/ aging of
red blood cells. CULLING is the destruction of senescent (aged) red blood cells by
the spleen.
Mechanisms:
1. Extravascular Hemolysis/ Macrophage-Mediated Hemolysis

This type of hemolysis happens within the reticuloendothelial system (SPLEEN)
when complement is not activated or incompletely activated. It accounts for
90% of red cell destruction and leads to increased unconjugated bilirubin &
urine/ fecal urobilinogen. It is the type of hemolysis seen in Rh hemolysis
2. Intravascular Hemolysis/ Fragmentation/ Intravascular Hemolysis

It happens within BLOOD VESSELS when the complement is completely
activated. It accounts for 10% destruction of aged red cell population and
leads to hemoglobinuria, decreased haptoglobin and hemopexin. It is the
type of hemolysis observed in ABO hemolysis.
Figure 2-16. Embden-Meyerhof Pathway
ACTIVITY 3 (Erythrocyte kinetics): Discuss
what happens to the red cell in each of the
given scenario: (3 points each)
1. Deficiency of integral proteins
2. Deficiency of peripheral proteins
3. Enzymatic deficiency in the EMP
4. Enzymatic deficiency in the PPP
5. Dysfunctionality in the Methemoglobin Reductase Pathway
6. Dysfunctionality in the Rapoport- Luebering Pathway
UNIT 4: LEUKOPOIESIS
Leukocytes are a heterogenous group that can be divided into two – the
granulocytes and agranulocytes. The granulocytes together with the monocytes share the
same lineage with the red cells (CFU-GEMM) while the lymphocytes have their own (CFU-
L).
A. GRANULOPOIESIS
Granulopoiesis pertains to the production and development of the three
granulocytes – the neutrophils, eosinophils and basophils. The maturation sequence is
almost similar for the three types of cells, except for the cytokines that influence
production and differentiation.
1. Neutrophil Development
Maturation Sequence (Fig. 2-17, Table 2-1)

I. Stem cell pool
a. Pluripotential hematopoietic stem cell
II. Mitotic pool
Progenitors:
a. CFU-GEMM (Common Myeloid Progenitor)
b. CFU-GM (Granulocyte-Macrophage Progenitor)
c. CFU-
G
Precursors:
d. Myeloblast
e. Promyelocytes
f. Myelocytes
III. Maturation pool
Precursors:
a. Metamyelocytes
b. Neutrophilic band
Figure 2-17. Neutrophil maturation sequence
Table 2-1. Characteristics of cells in the maturation sequence of granulocytes
Cell/ Stage Size N:C Nucleus Cytoplasm Cellular Activity
(um) ratio Shape Chromatin Nucle Staining Granules
oli
MYELOBLAST 14- 8:1 to Round Homogenous, 2 to 4 Slightly No Granules 0-3% of nucleated
20 4:1 to oval delicate, fine basophili cells in BM
euchromatin c * Classification:
Type I blasts:
No visible
granules
“Granular blasts”
Rare in normal
marrow
Type II blasts: < 20
visible primary or
azurophilic
granules
Type III blasts: >20

visible primary or
azurophilic
granules
PROMYELOCYTE 16- 3:1 to Round Heterochroma 1-3 Basophili Formation of 1-5% BM
25 2:1 to oval tin c PRIMARY/ Hof/ Paranuclear
Slightly coarse Azurophilic halo surrounding
granules the nucleus
MYELOCYTE 12- 1:1 Oval or Coarser and NONE Mixture Formation of 6-17% BM
18 round condensed of SECONDARY
(“Dawn of basophili / LAST STAGE
Neutrophilia”) c and Specific CAPABLE OF
acidophi granules MITOSIS
lic “Dawn of
Neutrophilia
”
METAMYELOCYTE 15- 1:1 KIDNEY- Coarse & NONE Beige/ Formation of 3-20% BM
18 SHAPED clumped salmon TERTIARY/
Gelatinase
granules
BAND/STAB 9-15 1:1 to Elongat Coarse & NONE Beige/ Continuous
1:2 e/ clumped salmon formation of
band tertiary
(C or S) granules
Formation of
SECRETORY
GRANULES
(vesicles)
Neutrophil Granules
Primary (Azurophilic) Granules
Formed during the promyelocyte stage
Last to be released (Exocytosis)
Contain:
 Myeloperoxidase
 Acid-β- glycerophosphate
 Cathepsins
 Defensins
 Elastase
 Proteinase-3
 Others
Secondary (Specific) Granules
Formed during myelocyte and metamyelocyte stages
Third to be released
Contain:
 β2- microglobulin
 Collagenase
 Gelatinase
 Lactoferrin
 Neutrophil gelatinase- associated lipocalin
 Transcobalamin I
 Others
Tertiary Granules
Formed during metamyelocyte and band stages
Second to be released
Contain:
 β2- microglobulin
 Collagenase
 Gelatinase
 Lysozyme
 Acetyltransferase
Secretory Granules (Secretory Vesicles)
Formed during the band and segmented neutrophil stages
First to be released (fuse to plasma membrane)
Contain (attached to the membrane):
 CD11b/ CD18
 Alkaline phosphatase
 Vesicle-associated membrane-2
 CD10, CD13, CD14, CD16
 Cytochrome b558
 Complement 1q receptor
 Complement receptor-1
Neutrophils are also known as polymorphonuclear cells (PMNs) or segmenters. They

are the cells that respond to bacterial infection. Their average size ranges from 9 to 15
microns. The nucleus presents with 2-5 lobes with highly condensed chromatin. The
cytoplasm will contain continuously forming pink to rose-violet secretory granules. They are
further characterized with the following:
- Normal values:
o Bone marrow: 7-30% of nucleated cell population
o Relative value in peripheral blood: 50-70% of WBCs
o Absolute value: 1.7-7.5 x 109/ L
- Neutrophil kinetics:
o Production: 0.9-1.0 x 109 cells/ kg per day
o Mitotic pool: 2.11 x 109 cells/ kg
o Maturation pool: 5.6 x 109 cells/ kg
o Once in the peripheral blood, neutrophils are divided randomly into a
circulating neutrophil pool (CNP) and a marginated neutrophil pool (MNP).
The ratio of CNP and MNP is roughly equal.
o Majority of the MNP are in the capillaries of the lungs.
- Transit time:
o HSC to myeloblast: 6 days
o Myeloblast to maturation pool: 4 to 6 days
o Neutrophil half-life in blood: 6-8 hours
o It takes about 14 days from the blast stage to the release of
mature granulocytes.
Neutrophil Function:
a. Phagocytosis
Chemical signals from damaged cells leading to chemotaxis  Margination (sticking

to capillary endothelium)  Diapedesis  Recognition of pathogen Attachment (Toll-like
receptor of phagocyte attaching to PAMPs)  Ingestion  Pathogen in phagosome 
Formation of phagolysosome Digestion & killing
Digestion:
 Oxygen dependent
 Respiratory burst through the activation of NADPH oxidase. H2O2
and peroxidase are produced
 Oxygen independent
 The pH within the phagosome becomes alkaline and then
neutral, the pH at which digestive enzymes work.
b. Generation of neutrophil extracellular traps (NETs)
o NETs
 Nuclear & organelle membrane dissolves  DNA release  DNA +
cytoplasmic enzymes  Cell membrane ruptures  NET release
 Extracellular threadlike structures believed to represent chains
of nucleosomes from DNA
 Have enzymes from neutrophil granules
 Have been shown to be able to trap and kill gram-positive and gram-
negative bacteria as well as fungi
“NETosis”: NETs are generated at the time that neutrophils die as a result of
o
antibacterial activity
c. Secretory Function
 Transcobalamin I/R binder (needed for Vitamin B12 absorption)
 Variety of cytokines
2. Eosinophil Development
Maturation Sequence
I. Pluripotential hematopoietic stem cell
II. Progenitors:
a. CFU-GEMM (Common Myeloid Progenitor)
b. CFU-Eo
III. Precursors:
a. Myeloblast
b. Promyelocytes
c. Myelocytes
d. Metamyelocytes
e. Eosinophilic band
IV. Eosinophil
Precursors:
A. Eosinophilic myeloblasts
o Not fully characterized
B. Promyelocytes
o Cytochemical identification only
o PRIMARY GRANULE: Charcot-Leyden crystal protein
C. Myelocytes
o Similar to neutrophil myelocytes
o Large, pale, reddish-orange SECONDARY GRANULES
D. Metamyelocytes & Band Forms
o Resemble their neutrophil counterpart
o Formation of SECRETORY GRANULE/ VESICLE
o Two other organelles are also present: Lipid bodies and Small lysosomal
granules
Eosinophil Granules
Primary Granules
Formed during the promyelocyte stage
Contain:
 Charcot-Leyden crystals
Formed throughout remaining maturation stages
Contain:
 Major basic protein (core)
 Eosinophil cationic protein (matrix)
 Eosinophil- derived neurotoxin (matrix)
 Eosinophil peroxidase (matrix)
 Lysozyme (matrix)
 Catalase (core and matrix)
 β-Glucuronidase (core and matrix)
 Cathepsin D (core and matrix)
 Interleukins 2,4, and 5 (core)
 Interleukin- 6 (matrix)
 Granulocyte- and Macrophage colony-
stimulating factor (core)
 Others
Small Lysosomal Granules
 Acid phosphatase
 Arylsulfatase B
 Catalase
 Cytochrome b558
 Elastase
 Eosinophil cationic protein
Lipid Bodies
 Cyclooxygenase
 5-Lipoxygenase
 15-Lipoxygenase
 Leukotriene C4 synthase
 Eosinophil peroxidase
 Esterase
Storage Vesicles
Carry proteins from secondary granules to be released into
the extracellular medium
Eosinophil specific granules can also be classified as larger or smaller granules

Secondary (specific) granules
Larger granules
MAJOR BASIC PROTEIN, Acid hydrolase, Peroxidase, Phospholipase,
Cathepsin, Eosinophil cationic protein, Eosinophil-derived neurotoxin
Smaller granules
Arylsulfatase, Peroxidase, Acid phosphatase
Eosinophils have bilobed nucleus measuring around 9-15 microns. They possess
refractile, orange-red granules and are involved in allergic and parasitic infections.
Eosinophils are further characterized by the following:
- Normal values:
o Relative value: 1-3% of WBC in peripheral blood
o Absolute value: 0-0.3 x 109/ L
- Production of eosinophil from last myelocyte division: 3.5 days
- Eosinophil kinetics:
o Turnover of eosinophils: 2.2 x 108 cells/ kg
o Large storage pool: 9-14 x 108 cells/ kg
o Half-life in circulation: 18 hours
o Survival in tissues: 2-5 days (columnar epithelial cells of the respiratory,
genitourinary, and gastrointestinal tracts)
Eosinophil Function:
a. Eosinophil degranulation
i. Classical exocytosis
 Granules move to plasma membrane  Fuse with cell membrane 
Emptying of contents to extracellular fluid (ECF)
ii. Compound exocytosis
 Granules fuse together within eosinophils  Fuses with cell membrane
 Emptying to ECF
iii. Piecemeal degranulation
 Secretory vesicles remove specific CHONs from 20 granules 
Secretory vesicles migrate to plasma membrane  Emptying to ECF
b. Regulation of immune responses

o Eosinophils delete double-positive thymocytes, act as antigen-presenting
cells, promote proliferation of effector T cells, initiate Type 1 or Type 2
immune response and regulate mast cells.
 Major basic protein & other cytokines can trigger mast cell
degranulation
 Nerve growth factor is needed for mast cell survival
c. Indicator of parasitic infections

o MAJOR BASIC PROTEIN & Eosinophil cationic protein destroys tissue-invading
helminths and prevents reinfection
d. Hallmark of allergic disorders

o In the peripheral blood, eosinophil concentration correlates with severity of
disease. They secrete HISTAMINASE, IL-5 that function for airway inflammation
and mucosal cell damage, and eosinophil-derived fibrogenic growth
factors that function for airway remodeling.
3. Basophil Development
Maturation Sequence:
II. Progenitors:
a. CFU-GEMM (CMP)
b. CFU-Baso
III. Immature basophil
IV. Mature basophil
Basophil Granules

Formed throughout remaining maturation stages
Contain:
 Histamine
 Platelet-activating factor
 Leukotriene C4
 Interleukin-4
 Interleukin-13
 Vascular endothelial growth factor A
 Vascular endothelial growth factor B
 Chondroitin sulfates (e.g. Heparin)
Basophils possess unsegmented or bilobed nucleus with condensed chromatin. The

blue-black water-soluble granules almost obscure the nuclear material of the cell.
Basophils are further characterized by:
- Normal values
Relative value: 0-2%
Absolute value: 0-0.2 x 109/ L
- Basophil kinetics:
Poorly understood
Life span of 60 hours
- Basophil functions: (ALLERGIC OR HYPERSENSITIVITY REACTION)
Basophils possess surface IgE receptors. They regulate Th2 response (IL-4 & IL-
3), induce B cells to synthesize IgE, mediate allergic processes (production of
HISTAMINE, Granzymes B, retinoic acid) and promote angiogenesis (Vascular
endothelial growth factor production)
Mast cells are erroneously called tissue basophils. They are not true leukocytes; they are cells
from the BM that uses blood as transit system to gain access to tissues where they mature.
They function as effector cells in allergic reactions by stimulating IgE receptors and
inflammatory reactions by an IgE receptor-independent process. They can also act as antigen
presenting cells that induce Th2 differentiation. They are known for their anti-inflammatory
and
B. AGRANULOPOIESIS immunosuppressive functions.
1. MONOPOIESIS
Maturation sequence (Table 2-2):
II. Progenitors:
a. CFU-GEMM (CMP)
b. CFU-GM
c. CFU-M
III. Precursors:
a. Monoblasts
b. Promonocyte
IV. Monocyte
V. Tissue spaces: Macrophage
Table 2-2. Characteristics of cells in the maturation sequence of monocytes

Cell/ Stage Size N:C Nucleus Cytoplasm Cellular Activity
(um) ratio Shape Chromati Nucle Staining Granules
n oli
MONOBLAST 12-20 4:1 to Round to Delicate 1-2 Basophili No Granules
3:1 oval c
PROMONOCYTE 12-18 3:1 to Slightly Delicate >1 Blue-gray Formation of Carries out 2
2:1 indented or AZUROPHILIC mitotic divisions
folded GRANULES in 60 hours to
produce 8
monocytes
Can carry out 4

mitotic divisions
in 60 hours
under
increased
demand
MONOCYTE 15-20 2:1 to Oval or Looser NONE Blue-gray Many fine Enter tissues
1:1 round (Lace- azurophilic and mature to
KIDNEY/ like/ granules macrophages
LARGEST CELL IN HORSE-SHOE Stringy) having
PERIPHERAL May be GROUND-
BLOOD
folded, GLASS
showing APPEARANCE
brain-like (frosted)
convolutions
Macrophages are large cells ranging from 40-50 microns. They present with
oval, reticulated chromatin nucleus and a pale, frequently vacuolated cytoplasm. They are
either wandering, or fixed as to location. They are given names according to their tissue
location:
 Liver: Kupffer cells
 Lungs: Alveolar macrophages
 Brain: Microglia
 Skin: Langerhans cells
 Spleen: Splenic macrophages
 Bone: Osteoclasts
 Synovial membrane: Type A cell
 Kidneys: Mesangial cells
 Lymph nodes: Dendritic cells
Monocyte/ Macrophage Kinetics

- Promonocyte pool
o 6 x108 cells/ kg
 Produces 7 x106 monocytes/ kg per hour
- NO STORAGE POOL: MONOCYTES ARE IMMEDIATELY RELEASED
o Pools in PB circulation: Circulating pool and marginal pool
 Marginal pool is 3.5x more numerous
o Half-life: 8.4 hours
o Stays for 3 days
- Normal values (Monocytes)
o Relative value: 2-11%
o Absolute value: 0.1-1.3 x 109/ L
Monocytes and macrophages function for innate immunity. They perform

phagocytosis via toll-like and Fc receptors and are involved in inflammatory responses.
They produce nitric oxide which is cytotoxic against viruses, bacteria, fungi, protozoa,
helminths and tumor cells. They also have housekeeping functions by removing debris and
dead cells, maintenance of iron storage pool and synthesis of a wide variety of proteins.
2. LYMPHOPOIESIS
Table 2-3. Characteristics of cells in the maturation sequence of lymphocytes
Stage Cell Size (microns) Nucleus Cytoplasm
LYMPHOBLAST 10-18 Coarse chromatin No granules present
Round or oval Appears smooth
With 1-2 nucleoli Moderate to dark
blue
PROLYMPHOCYTE Maybe same size as More clumped Usually nongranular
lymphoblast or chromatin Moderate to dark
smaller Round or oval in shape blue
With 1- 2 nucleoli
MATURE LYMPHOCYTE
SMALL LYMPHOCYTE 8-10 Dense chromatin Very scanty
Round or oval in shape ROBIN’S EGG BLUE
No nucleoli visible
OCCUPIES MAJORITY
OF CELL AREA
MEDIUM LYMPHOCYTE 10-12 Chromatin not as dense More abundant than

as small lymphocyte that in small lymphocyte
Round or oval in shape Pale to moderate blue
No nucleoli visible
LARGE LYMPHOCYTE 12-16 Round or oval in shape Abundant

(Atypical Lymphocyte) No nucleoli visible Clear, very pale blue
Functionally, lymphocytes are classified into three. T lymphocytes which are
responsible for cellular-mediated immunity comprise 60 to 80% of the lymphocyte
population and last for 4 to 10 years. B lymphocytes which are involved in humoral-
mediated immunity comprise about 10 to 20% of the lymphocyte population and last for
3 to 4 years. Null lymphocytes which are capable of tumor host defense comprise only
10% of the lymphocytes in the body.
Atypical, reactive or plasmacytoid lymphocytes are cells that morphologically

resemble monocytes. They are seen in the peripheral blood as a result of viral infections. To
differentiate between an atypical lymphocyte and a monocyte, the following should be
noted:
o Sharp indentation of the cytoplasm by adjacent red cells is a feature of

the reactive lymphocyte and is not observed for monocytes. (For reactive
lymphocytes, their cell membrane often gives way for the neighboring red
blood cells)
o Reactive lymphocytes characteristically have an increased amount of dark
blue cytoplasm, whereas monocyte cytoplasm is usually a blue-gray color
o Lymphocytes lack the many fine granules that give monocytes a typical
“ground glass” appearance of the cytoplasm
Lymphocytes have a relative value of 18-42% and an absolute value of 1- 3.2 x 109/ L.
a. CD4+/ Thelper
o Th1: Against intracellular pathogens
o Th2: Against extracellular parasites; Induction of asthma and allergic diseases
o Th17: Against extracellular bacteria and fungi
b. CD8+/ Tcytotoxic
o Secretes granules containing granzymes and perforins
o Killing of target cells
c. Treg
o Regulate immune response
o Maintains self-tolerance
d. B lymphocytes
o Antibody production (Plasma cell)
o Antigen presentation to T cells
 Optimal CD4+ activation
o Production of cytokines (Regulates T cell and antigen presentation)
**Natural Killer Cells

- Large granular lymphocytes
- Granules that are peroxidase negative (Kills tumor cells and some virus-infected cells)
- Participates in innate immunity unlike other types of lymphocytes
EXPERIMENT 3 in the laboratory is on Hematopoiesis. Review all the principles

in the previous topic and proceed to answer the ELABORATE requirement.
SAINT LOUIS UNIVERSITY
SCHOOL OF NATURAL SCIENCES
DEPARTMENT OF MEDICAL LABORATORY SCIENCE
Name: _
Instructor:
Rating:
Date:
EXPERIMENT 3
HEMATOPOIESIS
OBJECTIVES
1. Describe the principles of blood cell maturation.

2. Describe the stages of erythrocyte development, the order of maturation and series,
and characteristics of each stage with variation.
3. List the stages of leukocyte development, the order of maturation and series, and
characteristics of each stage with variation.
4. Describe the stages of thrombocyte development, the order of maturation and series,
and characteristics of each stage.
5. Describe the different components and functions of blood.
MATERIALS
Hematology atlas
Charts
PROCEDURE
1. Listen very carefully as the instructor discusses the different stages in the development of
the cellular components of the blood.
2. Using hematology atlases and charts, trace the different stages in the development of each
series. Take note of the characteristics of the different cells.
DRAWINGS
1. Maturation of human blood cells

2. Maturation sequences:
a. Cell size in relation to maturation and division
b. Cell size and color of cytoplasm
c. Nuclear morphology and chromatin structure
3. Chart showing the composition of blood.
QUESTIONS FOR RESEARCH
1. What are the general rules/trends of maturation?

2. What are the different growth factors that affect hematopoiesis?
3. What are the possible conditions of the bone marrow that will not support hematopoiesis? Explain.
4. What is meant by shift to left? Explain.
ELABORATE ACTIVITY 4 (CELL LINEAGES): In a flow chart, present

the individual maturation sequence of EACH normal
white blood cell. (5 points per blood cell lineage)
Include the hematopoietic growth factors or cytokines
involved in the maturation sequence. (25 POINTS)
(ONE COUPON BOND PER CELL LINEAGE)

MODULE 2 QUIZ
A. MCQ: CHOOSE THE BEST ANSWER. (15 POINTS)
1. Nucleated RBCs in neonates’ peripheral blood are presumed to come from
what hematopoietic development stage?
A. Mesoblastic B. Hepatic C. Myeloid D. Spleen, Thymus, Lymph nodes
2. During the first trimester of fetal development, the primary site of blood cell
production is the:
A. Bone marrow B. Spleen C. Yolk sac D. Liver
3. As most blood cells mature, which of the following is characteristic?
A. Cell diameter increases
B.Nucleus to cytoplasm ratio (N:C) decreases
C. Nuclear chromatin becomes less condensed
D. Basophilia of the cytoplasm increases
4. Which one of the following organs is responsible for the maturation of T lymphocytes and
regulation of their expression of CD4 and CD8?
A. Thymus B. Liver C. Spleen D. Lymph nodes
5. Myeloproliferative red cell population in the bone marrow of uremic patients is caused
by:
A. infiltration of bone marrow by toxic waste products
B. decreased levels of circulating erythropoietin
C. defective globin synthesis
D. overcrowding of bone marrow space by increased myeloid precursors
6. Megaloblastic asynchronous development in the bone marrow indicates which one
of the following?
A. Proliferation of erythrocyte precursors B. Impaired synthesis of DNA
C. Inadequate production of erythropoietin D. Immature release of reticulocytes
7. For neutrophils to defend against infection, they must be capable of all of the following,
except:
A. mobility B. cell division C. chemotaxis D. adhesion
8. What is the expected response of the body of a patient suffering from severe anemia
due to myelofibrosis?
A. Extramedullary hematopoiesis in the liver and spleen
B.Decreased production of erythropoietin by the kidney
C. Increased apoptosis of erythrocyte progenitor cells
D. Increase the proportion of yellow marrow in the long bones
9. Vasoactive compounds are principal components of the granules of:
A. monocytes B. basophils C. eosinophils D. neutrophils
10.The metabolic pathway that facilitates the release of oxygen from hemoglobin to the
tissues is:
A. Methemoglobin reductase. B. Rapoport-Luebering shunt.
C. Hexose-monophosphate shunt. D. Embden-Meyerhoff pathway.
11. The best source of active bone marrow from a 25-year-old male patient would be:
A. Iliac crest B. Femur C. Distal radius D. Tibia
12.Which of the following hemokines is involved in the early phases of differentiation

of hematopoietic stem cells?
A. IL-2 B. IL-8 C. EPO D. FLT3 ligand
For Nos. 13 – 15:

WRITE: A, if statement A is true and B is false;
B, if statement B is true and A is false;
C, if both statements are true;
D, if both statements are false.
13. A. In the red cell maturation sequence, the acidophilic erythroblast is the last cell
capable of mitosis.
B. It is also in this stage that the highly-condensed nucleus is ejected.
14. A. Mesenchymal hematopoiesis ends 6 months after conception.

B. Hepatic hematopoiesis begins on the third month following conception.
15. A. In granulocyte production, mitosis occurs in the proliferative pool

B. Differentiation of granulocytes occur in the post-mitotic pool.
B. MATCHING TYPE. Match the following Growth Factors with their Primary Target (10
POINTS)
A Growth Factors B Target Cells
1. G-CSF - D A. Basophils
2. Il-5 - G B. Megakaryocytes
3. Transforming Growth Factor B - A C. Erythroid Precursors
4. IL-6 - I D. Neutrophils
5. IL-11 - B E. Primitive Progenitor Cells
6. TPO - B F. Monocytes
7. Kit-Ligand - E G. T-cells
8. IL-9 – G H. Adipocytes
9. FL – E I. Malignant Plasma Cells
10.EPO - C J. Macrocytes

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MODULE 2

Hematopoiesis (Fig. 2-1) is the process of cellular

Figure 2-1. Overview of Cellular Maturation

1. Mesoblastic Phase (Fig. 2-2)

Figure 2-2. Embryonic hematopoiesis

Figure 2-2. Phases of Hematopoiesis

Figure 2-4. Sites of Hematopoiesis

Adult Hematopoietic Tissues

A. Primary lymphoid tissues

1. BONE MARROW (BM)

ACTIVITY 1 (MYELOID HEMATOPOIESIS): From the illustration of the skeletal system

C. Trilaminar sinus wall

Figure 2-7. Histology of the red bone marrow

Bone Marrow Specimens

Collection is oftentimes carried out to observe the patient’s myeloid-to-erythroid ratio

- Parts (Fig. 2-8, right):

B. Secondary lymphoid tissues

Figure 2-9. Structure of the lymph node

 Paracortex (T-cell area)

Figure 2-10. Structure of the Spleen

STEM CELL THEORY

STEM CELL DIVISION

S phase (8 hours) Replication of DNA and chromosomes

Models of Stem Cell Fate

CYTOKINES AND GROWTH FACTORS

Colony-stimulating Factors (CSF)

Growth factors can be classified according to the part of the development

ii. FLT3 ligand

Figure 2-12. Significant changes in blood cell maturation

ACTIVITY 2 (CELL MATURATION): According to the general rules (CAP Guidelines) of

**Three Erythroid Precursor Nomenclature Systems

Image from: https://www.google.com/search?q=eryhtropoiesis&tbm=isch&ved=2ahUKEwi6prCNlZXrAhUFHKYKHVFmC6sQ2-cCegQIABAA&oq=eryhtropoiesis&gs_lcp=CgNpbWcQAzIGCAAQChAYOgQIABBDOgUIABCxAzoCCABQu-

THE RBC MEMBRANE

Figure 2-14. Structure of the red cell membrane

The glycocalyx is an extensive sugary coat made up of the carbohydrate portions of

1. Embden-Meyerhof Pathway (EMP)

 ANAEROBIC PATHWAY/ ANAEROBIC GLYCOLYSIS

2. Hexose Monophosphate Pathway/ Pentose Phosphate Shunt

3. Methemoglobin Reductase Pathway

4. Rapoport- Leubering Pathway

1. Extravascular Hemolysis/ Macrophage-Mediated Hemolysis

2. Intravascular Hemolysis/ Fragmentation/ Intravascular Hemolysis

1. Deficiency of integral proteins

2. Deficiency of peripheral proteins

3. Enzymatic deficiency in the EMP

4. Enzymatic deficiency in the PPP

5. Dysfunctionality in the Methemoglobin Reductase Pathway

6. Dysfunctionality in the Rapoport- Luebering Pathway

Maturation Sequence (Fig. 2-17, Table 2-1)

Figure 2-17. Neutrophil maturation sequence

Type III blasts: >20

Neutrophils are also known as polymorphonuclear cells (PMNs) or segmenters. They

Chemical signals from damaged cells leading to chemotaxis  Margination (sticking

Eosinophil specific granules can also be classified as larger or smaller granules

b. Regulation of immune responses

c. Indicator of parasitic infections

d. Hallmark of allergic disorders

Secondary (Specific) Granules

Basophils possess unsegmented or bilobed nucleus with condensed chromatin. The

Table 2-2. Characteristics of cells in the maturation sequence of monocytes

Can carry out 4

Monocyte/ Macrophage Kinetics

Monocytes and macrophages function for innate immunity. They perform

MEDIUM LYMPHOCYTE 10-12 Chromatin not as dense More abundant than