Unit 2

HEMATOPOIESIS

Introduction

Hemopoiesis/hematopoiesis is a continuous, regulated process of

blood cell production that includes cell renewal, proliferation, differentiation,
and maturation. These processes result in the formation, development, and
specialization of all of the functional blood cells that are released from the
bone marrow to the circulation.

It refers to the formation and development of all types of blood cells

from their parental precursors. In postnatal life in humans, erythrocytes,
granulocytes, monocytes, and platelets are normally produced only in the
bone marrow. Lymphocytes are produced in the secondary lymphoid
organs, as well as in the bone marrow and thymus gland. A hypothetical
scheme of hemopoiesis based on a monophyletic theory is accepted by
many hematologists. According to this theory, the main blood cell groups
including the red blood cells, white blood cells and platelets are derived
from a pluripotent stem cell.

Learning Outcomes

At the end of this unit, students will be able to:

1. Define and explain the theories and principles involved in the

formation of red blood cells, white blood cells and platelets.
2. Describe the evolution and formation of blood cells from embryo to
fetus to adult, including anatomic sites and cells produced.
3. Describe general morphologic changes that occur during blood
cell maturation.
4. Discuss the process of Bone Marrow examination .

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Topic 1: The Origin, Formation and Development of Blood Cells

Learning Objectives

At the end of this topic, students will be able to:

1. Discuss the general processes involved in the formation of red blood cells,
white blood cells, and platelets.
2. Discuss the importance of certain cytokines in the formation of blood cells.
3. Enumerate the three phases of hematopoiesis.

Activating Prior Learning

Do you have an idea how are blood cells are formed, which organ is involved and
have you ever thought if there are nutritional requirements needed for it’s development?

Presentation of Contents

Terminology:

1. Pluripotent hematopoietic stem cell (HSC)

Undifferentiated cell producing blood cells of all lineages, capable of self-renewal
2. Multipotent HSC
Undifferentiated cell producing cells of multiple lineages, limited self-renewal (e.g.,
myeloid SC, lymphoid SC)
3. Committed progenitor - undifferentiated cell capable of producing cells of one lineage,
colony forming units (CFUs) (e.g., erythroid CFU, granulocyte-macrophage CFU)
4. Hematopoietic stem cell (HSC): defined functionally as the one cell capable of self-
renewing and generating all cell types in the blood.
5. Myeloid- general term used to denote granulocytic cells and their precursors, including
basophils, eosinophils, and neutrophils. Lymphoid and erythroid cell lines are excluded,
and most morphologists also exclude the monocytic and megakaryocytic cell lines.
6. Lymphoid- resembling or pertaining to lymph or tissue and cells of the lymphoid
system.

Types of Cellular Element

1. Erythrocytes- Red Blood Cells
2. Leukocytes- White Blood Cells
a. Neutrophils- Segmenters
b. Eosinophils

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 c. Basophils
d. Lymphocytes
e. Monocytes
3. Thrombocytes- Platelets

Hematopoiesis
Figure 1: Hematopoiesis 2 Main Branches: Myeloid and Lymphoid

STAGES OF HEMATOPOIESIS

A. PRENATAL HEMATOPOIESIS
I. Mesoblastic Period
Yolk sac - chief site
2nd week of fetal life, formation of blood islands in yolk sac (mesodermal extraembryonic
layer), aggregation of primitive cells
9th week of fetal life, development of primitive erythroblast
o Primitive Erythroblast – produces Hb

Gower I
Gower II three embryonic forms of Hb
Portland

II. Hepatic Period

Starts on the 2nd month
Liver - chief site; 3rd month of fetal life
SIGNIFICANT CONTRIBUTIONS OF OTHER ORGANS:
a) Thymus – T cell production
b) Kidney – B cell
c) Spleen-active in:
i. erythropoiesis- until the end of normal gestation (splenic)

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 ii. myelopoiesis – but becomes minimal by the 5th month
iii. Lymphopoiesis – lifetime
a) Lymph nodes
NRBC’s , Granulocytes, Monocytes, Lymphocytes and Megakaryocytes
Hb A
Hb A2 hemoglobins
Hb F

III. Medullary/ Myeloid Period

Red bone marrow - chief site
Starts on the 5th month of fetal life
Development of cells increases at birth
Hb A1 - begins to appear and gradually increases in concentration
EPO, G-CSF, GM-CSF, Hb F, Hb A2

B. ADULT HEMATOPOIESIS
I. Bone Marrow- is the only site of erythropoiesis, myelopoiesis and thrombopoiesis
1st few years of life – a delicate balance exists between developing bone marrow space and
the developing infant’s need for blood cells and the liver or spleen remains available bec.
of its hematopoietic capability
4th year of life – rate of bone marrow growth exceeds the need for blood cells therefore,
active marrow sites are replaced with areas of fatty reserve and fat first develops in the long
bones
18th year of life – the only active hematopoietic sites are in the pelvis, vertabrae, ribs,
sternum, skull and proximal extremities of the long bones
2 Major Components
i. Red Marrow- hematopoietically active marrow consisting of the developing blood
cells and their progenitors
– Site of blood cell development . It comprises approximately 50% of the marrow
cavity space.
The remaining 50% of the space is occupied by fat and is known as yellow marrow.
The ratio of red marrow to yellow marrow is an indirect representation of marrow
activity, and is expressed as marrow cellularity.
ii. Yellow Marrow- hematopoietically inactive marrow composed primarily of adipocytes
(fat cells), with undifferentiated mesenchymal cells and macrophages.
Retrogression- process of replacing the active marrow by adipocytes (yellow marrow) during
development
Develops in the embryo by the hollowing out of the skeletal bones forming a central cavity
In this cavity develops a primitive, undifferentiated cell known as a hemocytoblast, or stem
cell
ALL blood formed elements ultimately develop from this undifferentiated precursor

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Graphic illustration of the arrangement of the extravascular area in hematopoietic
tissue

ADIPOCYTES

HEMATOPOIETIC TISSUE

BONE

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 THE NORMAL BONE MARROW

- Full of developing precursor

cells in all stages of
maturation

- Released into blood at

Maturity

- ONLY mature cells are

released normally

- Blast - first recognizable

precursor in each cell
line

- Marrow recedes during

development of the individual,
and in the adult occupies only
the support skeleton and the
proximal regions of the long
bones

II. EXTRAMEDULLARY HEMATOPOIESIS

Blood cell production in hematopoietic tissue other than bone marrow.
Occurs when hyperplasia (increase in number of cells per volume of tissue) of
marrow cannot meet physiologic blood needs of tissue.
Principally occurs in liver and spleen (just like in fetus) lymph nodes and
thymus
1. LIVER
Cellular Functions:
Synthesizes various transport proteins
Stores essential minerals and vitamins that are used in DNA and RNA synthesis
Conjugation of bilirubin Hb degradation
Transports bilirubin to the small intestines for excretion
Kupffer cells – macrophages in liver cells

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 2. SPLEEN
Reticuloendothelial organ
Powerhouse of prominent red blood cell activity such as filtration,production
and cellular immunity.
Blood-filled organ that consists:
o Red pulp = red blood cell filtration
o White pulp = lymphocyte processing
o Marginal zones = stores wbc’s & platelets
Primary functions - lymphopoiesis, phagocytosis
Located in the left side of the abdomen just below the diaphragm and behind
the fundus of the stomach
Largest structure of the lymphoid system

3. LYMPH NODES
Oval structures distributed throughout the body connected by lymphatic vessels which
carry a fluid called lymph.
Act as filters to remove foreign blood contaminants. Extremely important part of the
body’s infection defense
Contain many phagocytic cells and lymphocytes
Immature lymphocytes produced in the bone marrow collect and mature in the tissues of
the nodes

Histologic structure of a normal lymph node.

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 4. THYMUS
- ductless gland located above the heart that plays a role in immunity
Thymus Gland
o responsible for normal development of some of the lymphocytes
o located in the neck
o maximum development in childhood, atrophies with age
Bursa Fabricus
o found in birds with possible analogous tissue in man. Responsible for normal antibody
production

Note the differences in size of the thymus of the infant and the adult

 STEM CELL THEORY OF HEMATOPOIESIS

- All cells are derived from a pool of stem cells that are self-renewing
Pluripotential & multipotential stem cells give rise to committed stem cells for each cell
line
Committed stem cells have receptors for specific growth factors
Respond to stimulation by division & maturation (precursor cell stages) into end-stage
cells

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Kierszenbaum, Fig. 6-16; See Ross 5th Ed Table 10.4

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 CYTOKINES
- Secreted proteins that bind to cell surface receptors and transduce signals leading to the
differentiation or proliferation of cells.
- Stimulates a specific lineage of cell
- Control replication and clonal or lineage selection & are responsible for maturation rate &
growth inhibition of stem cells
- include colony-stimulating factors, interferons, interleukins, lymphokines, monokines
- such as erythropoietin, G-CSF, G-CSF, interleukins, etc

 REGULATION OF HEMATOPOIESIS
- Erythropoietin (EPO) - major regulator of erythropoiesis, stimulates erythroid CFU cells
and proerythroblasts
r-HuEPO - human recombinant erythropoietin
- pharmaceutically available product
- Thrombopoietin (TPO) - increases platelet production, stimulates megakaryocyte CFU
cells
- Granulocyte CSF (G-CSF) - increases production of neutrophils, stimulates granulocyte-
macrophage CFU cells
- Granulocyte-macrophage CSF (GM-CSF) - increases macrophage production,
stimulates granulocyte-macrophage CFU cells
- Interleukins - stimulate B- and T-cell formation, function together with G-CSF and GM-
CSF
- KITLG & FLT3L – Early – acting growth factors

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 Terminology
- Pluripotent hematopoietic stem cell (HSC)
Undifferentiated cell producing blood cells of all lineages, capable of self-renewal
- Multipotent HSC
Undifferentiated cell producing cells of multiple lineages, limited self-renewal (e.g.,
myeloid SC, lymphoid SC)
- Committed progenitor - undifferentiated cell capable of producing cells of one lineage,
colony forming units (CFUs) (e.g., erythroid CFU, granulocyte-macrophage CFU)

 PRIMITIVE CELLS:
- Hemocytoblasts (hematopoietic stem cells)
Give rise to all formed elements
Hormones and growth factors push the cell toward a specific pathway of blood
cell development

A. ERYTHROPOIESIS:
- A hemocytoblast is transformed into a proerythroblast
- Proerythroblasts develop into early erythroblasts
- The developmental pathway consists of three phases
1. ribosome synthesis in early erythroblasts
2. Hb accumulation in late erythroblasts and normoblasts
3. ejection of the nucleus from normoblasts and formation of reticulocytes
- Reticulocytes then become mature erythrocytes

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CD 34 – hematopoietic stem cell marker

THE LIFE CYCLE OF A RED BLOOD CELL:

a. Kidneys respond to a lower than normal oxygen concentration in the blood by releasing
the hormone erythropoietin.
b. Erythropoietin travels to the red bone marrow and stimulates an increase in the
production of red blood cells (RBCs).
c. The red bone marrow manufactures RBCs from stem cells that live inside the marrow.
d. RBCs squeeze through blood vessel membranes to enter the circulation.
e. The heart and lungs work to supply continuous movement and oxygenation of RBCs.
f. Damaged or old RBCs are destroyed primarily by the spleen.

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 NUTRITIONAL REQUIREMENTS:
- Proteins, lipids, and carbohydrates
- Iron, vitamin B12, and folic acid
- The body stores iron in Hb (65%)
Intracellular iron is stored in protein-iron complexes such as ferritin and
hemosiderin
Circulating iron is loosely bound to the transport protein transferrin

 ORGANS INVOLVED IN ERYTHROPOISESIS:

 HORMONAL CONTROL OF ERYTHROPOIESIS

- Erythropoietin (EPO)
Direct stimulus for erythropoiesis
Released by the kidneys in response to hypoxia
- Causes of hypoxia
Hemorrhage or increased RBC destruction reduces RBC numbers
Insufficient hemoglobin (e.g., iron deficiency)
Reduced availability of O2 (e.g., high altitudes)
- Effects of EPO
More rapid maturation of committed bone marrow cells
Increased circulating reticulocyte count in 1–2 days
- Testosterone also enhances EPO production, resulting in higher RBC counts in males

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 ERYTHROCYTE DESTRUCTION
As the red cell ages, changes occur that make it susceptible to destruction
Alteration in the membrane integrity takes place
Loss of sialic acid and lipids, decreased ATP and increased Calcium have been
implicated in the aging process
At 120 days the erythrocytes are recognized as abnormal and are removed by
phagocytic cell in the RES
As the cell ages it is depleted of glucose and their surface area decreases
The spleen recognizes abnormalities in the cell and sequester it for removal

FORMATION & DESTRUCTION OF RBCS

LEUKOPOIEIS
- PRODUCTION OF LEUKOCYTES
- stimulated by interleukins and colony-stimulating factors (CSFs)
Interleukins are numbered (e.g., IL-1, IL-2), whereas CSFs are named for the
WBCs they stimulate (e.g., granulocyte- CSF - stimulates granulocytes)
- Macrophages and T cells are the most important sources of cytokines

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FORMATION OF LEUKOCYTES:
- All leukocytes originate from hemocytoblasts
- Hemocytoblasts differentiate into myeloid stem cells and lymphoid stem cells
- Myeloid stem cells become eosinophilic, basophilic and neutrophilic myeloblasts or
monoblasts
- Lymphoid stem cells become lymphoblasts
- The myeloblasts develop into eosinophils, neutrophils, and basophils
- Monoblasts develop into monocytes
- Lymphoblasts develop into lymphocytes

 MYELOID LINEAGE

- Myeloid lineage gives rise to granulocytes, and monocytes (and macrophages), which are
not granulocytes. Diagram does not show changes in cell and nuclear morphologies.
Band stage is not shown above.
- 14 days - Maturation period of most granulocytes from blast stage until to the release of
mature granulocytes into the peripheral blood

Read: Table 7.2 Rodak – Selected Cytokines, Characteristics, Current & Potential Therapeutic
Applications pg. 90 – 91

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B.

C. LYMPHOPOIESIS:
- Lymphocytes are derived from committed stem cells that originate from pluripotent stem
cell.
- Early lymphoid cells further differentiates into B & T lymphocytes.

 LYMPHOID LINEAGE
- Lymphoid stem cell gives rise to T-lymphocyte
and B- lymphocyte lineages
- T-cell maturation - thymus
- B-cell maturation - bone marrow
- Plasma cells - present in marrow,
lymphatic tissue, connective tissue

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 LYMPHOCYTIC MATURATION SEQUENCE
- LYMPHOBLAST
- PROLYMPHOCYTE
- LYMPHOCYTE

 LYMPHOPOIESIS
- Immunologically competent cells
- Primary lymphoid organs
Bone marrow
Thymus
- Secondary lymphoid organs
Lymph nodes
Spleen
Peyer’s patches
Tonsils
- Lymphocytes
Originate in the BM, thymus gland & the lymphatic system
B lymphocytes
 represent 10 – 20% of the total lymphocyte count
 short – lived ( 3- 4 days)
 responsible for humoral-mediated immunity (plasma cells = antibody
production)
T lymphocytes
 represent 60 – 80% of the total lymphocyte count
 long – lived (4-10 years)
 essential in cellular – mediated immunity (T helper &T
cytotoxic/suppressor cell)
NK cells
 a lymphocyte in defense against to certain tumor cells and
virus-infected cells without the stimulation of antigens, and kill them by
the insertion of granules containing perforin.
 tumor – host defense
 Null cells
 10% of the lymphoid population

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D. MONOPOIESIS
- Development of the monocyte
- Stages in the monocytic development are:
Monoblast
Promonocyte
Monocyte 

- Distribution - blood and CT (as transient or wandering cells)

- Function - immune protection, movement (cell motility)
- Diapedesis - movement or passage of white blood cells through intact capillary walls into
surrounding body tissue. - also called migration
- Chemotaxis - movement of cells in response to the influence of chemical stimulation
- Chemokines - chemotactic cytokines with signaling proteins secreted by cells induced to
direct chemotaxis in nearby responsive cells

E. MEGAKARYOPOIESIS
 THROMBOPOIESIS: GENESIS OF PLATELETS
- Production of platelets directly from the cytoplasm of metamegakaryocyte
- The stem cell for platelets is the hemocytoblast
- The sequential developmental pathway is as shown:

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 STAGES OF MATURATION OF MEGAKARYOCYTES:
- MEGAKARYOBLAST
- PROMEGAKARYOCYTE
- GRANULAR MEGAKARYOCYTE
- MATURE MEGAKARYOCYTE (METAMEGAKARYOCYTE)
In differentiating the maturation stages of the megakaryocytic cells, emphasis should be
placed on the cytoplasmic appearance rather than the number of nuclei or chromaton
structure.(Steininger)

 PLATELETS
- Small fragments of metamegakaryocytes
- Formation is regulated by thrombopoietin
- Cytoplasm: Light blue to purple outer region, very granular
Chromomere = granular & located centrally
= granulomere
= tubules, dense granules and other
= components
Hyalomere = surrounds the chromomere, nongranular & clear to light blue
= microtubules & filaments
- Granules contain serotonin, Ca2+, enzymes, ADP, and platelet-derived growth factor
(PDGF)
- Development occurs via ENDOMITOSIS (megakaryopoiesis)
Neutrophils- 40-70
Endomitosis = Nuclear division without cytoplasmic division
- Life span: 9 – 12 days (4 – 9 days)
- Size: 1 - 4 um in diameter
- Disc shape or irregular in shape
- Functions:
maintenance of hemostasis
blood clotting
wound repair, etc.

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NORMAL CELLULAR MATURATION OF BLOOD CELLS:
1. Cytoplasmic Maturation
- Immature cell: intensely basophilic (blue) because of high RNA content. Loss of
blue as cell matures.
- Granules may appear as cell matures. 1st granules are nonspecific. Take on
specific characteristics as they mature.
- Relative amount of cytoplasm increases as cell matures.

2. Nuclear Maturation
- Immature nucleus is round or oval and large in proportion to rest of cell.
- As cell matures, decreases in size and takes on varying shapes.
- Nuclear chromatin changes from delicate and fine to coarse and clumped.
- Staining properties change from reddish purple to bluish purple.
- Nucleoli gradually disappear.

3. Cell Size
- As cell matures, becomes smaller in size

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