Ch13 Hematopoiesis

Prepared and presented by:

Dr. Salamah Alwahsh Histology II (19021)
‫برنامج الطب البشري‬
‫كلية الطب والعلوم الصحية‬
‫جامعة بولتكنيك فلسطين‬

06.09.2021

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 Hematopoiesis
• Mature blood cells have a relatively short life span, and must be continuously
replaced with progeny of stem cells produced in Hematopoiesis

Hematopoietic tissues
• In the early embryo these blood
cells arise in the

• yolk sac mesoderm → earliest

stage of embryogenesis

• The liver and spleen →

temporarily (starts in the 2nd
trimester)

• The bone marrow (medullary

cavities) of bones starts in the
2
3rd trimester
Pluripotential Hematopoietic stem cells

• Stem cells are pluripotential cells that are capable of self

renewal and division
 Some of their daughter cells form specific irreversibly
differentiated cell types (committed progenitor)
• Committed/progenitor cells - parent or ancestor cells that differentiate into one
cell line.
 other daughter cells remain stem cells to maintain a
constant number of pluripotential stem cells
Self-renewal - property of regenerating the same cells.

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Pluripotent hematopoietic stem cells

These cells can proliferate to produce all blood cell types :

 one cell lineage will form lymphocytes → lymphoid cells
(migrate from the bone marrow to the thymus, spleen,
lymph nodes )
 another cell lineage will form the myeloid cells that
develop in bone marrow (granulocytes, monocytes,
erythrocytes, and megakaryocytes)
• Pluripotential cell - cell that differentiates into many different
cell lines. It has the potential to self-renew, proliferate, and
differentiate into erythrocytic, myelocytic, monocytic,
lymphocytic, and megakaryocytic blood cell lineages

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Progenitor and Precursor Cells
• Hematopoiesis is the result of simultaneous, continuous
proliferation and differentiation of cells derived from stem
cells whose potentiality is reduced as differentiation
progresses

• The proliferating stem cells form daughter cells with

reduced potentiality:
 Like stem cells are not morphologically distinguished

 they resemble large lymphocytes

 Stem cells divide at a rate sufficient to maintain their relatively small

population. The rate is accelerated in progenitor and precursor cells

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 progenitor cells can divide and produce both progenitor and
precursor cells
In precursor cells (blasts) :
 The morphological characteristics differentiate for the first time
 They can produce only mature blood cells
 The progenitor cells for blood cells are often called colony-
forming units (CFUs), because they give rise to colonies of
only one cell type when cultured in vitro or injected into a
spleen

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Erythropoiesis - formation and maturation of erythrocytes in the bone
marrow. It is under the influence of the hematopoietic growth factor
erythropoietin.
Erythropoietin - hormone secreted by the kidney that regulates erythrocyte
production by stimulating the stem cells of the bone marrow to mature into
erythrocytes. Its primary effect is on the committed stem cell CFU-E.
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Microenivronment (niche) and cell differentiation
• Hemopoiesis depends on a microenvironment, or niche, with specific
endocrine, paracrine, and juxtacrine factors

• These requirements are provided largely by the local cells and

extracellular matrix (ECM) of the hemopoietic organs that together
create the niches in which stem cells are maintained and progenitor
cells develop

• Hemopoietic growth factors (colony-stimulating factors (CSF)) or

cytokines, are glycoproteins that stimulate proliferation of progenitor and
precursor cells and promote cell differentiation and maturation within
specific lineages

 stimulate proliferation (mitogenic activity) of immature cells

 support the differentiation of maturing cells
 enhance the functions of mature cells
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+ differentiation

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 Interferon: paracrine factors from leukocytes and
virus-infected cells that signal NK cells to kill the
infected cells and adjacent cells to resist viral
infection (minimise the replication of the virus)

Depolarisation of monocytes
to become anti-inflammatory

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Cytokines
• Cells in the immune system communicate with each other primarily via
cytokines to coordinate defensive measures
• Major responses induced in target cells by such factors are the following:
• Directed cell movements, or chemotaxis, toward and cell accumulation at sites of
inflammation, for example, during diapedesis. Cytokines producing this effect are also
called chemokines
• Increased mitotic activity in certain leukocytes, both locally and in the bone marrow
• Stimulation or suppression of lymphocyte activities in adaptive immunity. A group of
cytokines with such effects were named interleukins because they were thought to be
produced by and to target only leukocytes
• Stimulated phagocytosis or directed cell killing by innate immune cells
• Most cytokines have multiple target cells in which they exert several effects. Some are
produced by and target cells besides immune cells, including endothelial cells, certain
autonomic neurons, and cells of the endocrine system

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Differentiation of pluripotential stem cells during hematopoiesis 15
The rare pluripotent
hemopoietic stem cells divide
slowly, maintain their own
population, and give rise to 2
major cell lineages of
progenitor cells: the myeloid
and lymphoid stem cells.
The myeloid lineage includes
precursor cells (blasts) for
erythropoiesis,
thrombopoiesis,
granulopoiesis, and
monocytopoiesis, all in the
bone marrow
The lymphoid lineage forms B
and T lymphocytes and related
cells called natural killer cells,
with the later differentiative
stages occurring in lymphoid
organs.
Erythropoietin (EPO), colony-
stimulating factors (CSF),
cytokines, and growth factors
promote growth and differentiation
throughout these developmental
processes. 16
As the process of hematopoiesis takes place

 both the potential for differentiation and the self-renewing

capacity gradually decrease

 the mitotic response to growth factors gradually increase,

attaining its maximum in the middle of the process

 from that point on, mitotic activity decreases, morphological

characteristics and functional activity develop, and the mature
cells are formed

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Bone marrow
• Bone marrow is found in the medullary canals of long bones and
in the small cavities of cancellous bone, with two types based on
their appearance at gross examination:
• Blood-forming red bone marrow, whose color is produced by an
abundance of blood and hemopoietic cells, and
• Yellow bone marrow, which is filled with adipocytes that exclude
most hemopoietic cells
• In the newborn, all the bone marrow is red. As the child grows,
most of the bone marrow changes gradually into the yellow
variety
• Under certain conditions, such as sever bleeding or hypoxia,
yellow bone marrow is replaced by the red bone marrow 18
Distribution of red marrow (hematopoietic active) in the adult . This type
of bone marrow tends to be located in cancellous bone tissue .
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Red bone marrow
It is composed of :
 stroma: It is a three – dimensional meshwork of reticular cells
and a delicate web of reticular fibers containing hematopoietic
cells and macrophages
It contains collagen types I and III, fibronectin, laminin, and
proteoglycans
The sinusoidal capillaries
An external discontinuous layer of reticular cells and a loose
net of reticular fibers reinforce the sinusoidal capillaries
 hematopoietic cords .

 sinusoidal capillaries . .

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Section of active red bone marrow
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Maturation of Erythrocytes

Basic process in maturation:

• Synthesis of hemoglobin

• Formation of an enucleated, biconcave, small

corpuscle, the erythrocyte

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Major changes during maturation:

• Cell volume decrease

• The nucleoli become invisible in the light microscope
• Chromatin become more dense, the nucleus presents
a pyknotic appearance and extrude from the cell
• Gradual decrease in the number of polyribosomes
• Simultaneous increase in the amount of hemoglobin
• Gradual disappearance of mitochondria and other
organelles
•The production of RBCs required erythropoietin hormone
and iron, folic acid, and Vitamin B12
 Differentiation
• Proerythroblast: large cell, loose lacy chromatin, visible nucleoli,
basophilic cytoplasm.

• Basophilic erythroblast: strongly basophilic cytoplasm,

condensed nucleus, no visible nucleolus.

• Polychromatophilic erythroblast: condensed nucleus, acidophilic

cytoplasm.

• Orthochromatophilic erythroblast: no nucleus , thin layer of

cytoplasm, small number of ribosomes

• Reticulocyte: loose polyribosome and becomes a mature

erythrocyte.

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Pyknotic - pertaining to
degeneration of the nucleus of the
cell in which the chromatin
condenses to a solid, structureless
mass and shrinks.

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Medical application
• Red bone marrow also contains stem cells that can produce other
tissues in addition to blood cells
• These pluripotent cells may make it possible to generate specialized
cells that are not rejected by the body because they are produced
from stem cells from the marrow of the same patient
• The procedure is to collect bone marrow stem cells, cultivate them in
appropriate medium for their differentiation to the cell type needed for
transplant, and then use the resulting cells to replace defective cells
• These studies in regenerative medicine are at early stages, but
results with animal models are promising

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Granulopoiesis
Maturation of granulocytes involve synthesis of a number of
proteins that are packed into two organelles:

• Azurophilic granules: the first stage, and contain enzymes of

the lysosomal system (stain with basic dyes)

• Specific granules: the second stage, contain different proteins

in each type of granulocytes and are used for their activities.

• A shift in gene expression permitting:

Neutrophils to specialize in bacterial destruction
Esoinophils and basophils in inflammation regulation

• Nonspecific granules - large, blue-black granules found in promyelocytes.

• The granules have a phospholipid membrane and stain positive for peroxidase

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 Maturation of granulocytes
• Myeloblast: finely dispersed chromatin, visible nucleoli.
• Promyelocyte: basophilic cytoplasm, azurophilic granules
( lysosomal enzymes and myeloperoxidase), gives rise to
the three types of granulocyte

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 Kinetics of neutrophil production
• The total time taken for a myeloblast to emerge as a
mature neutrophil in the circulation is 11 days
• under normal circumstances 5 mitotic divisions occur
in (myeloblast, promyelocyte , and neutrophil myelocyte
stages of development) .

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The common features between the different
compartments
Between the marginating and circulating compartments :

1) About equal size .

2) There is a constant interchange of cells between them.

3) The half life of a neutrophil in these compartments is 6-7 h

The medullary formation and storage compartments together

are about 10 times as large as circulating and marginating
compartments .

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Mitosis

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_ Neutrophils and other granulocytes enter the connective
tissue by passing through INTERCELLULAR JUNCTIONS
found between endothelial cells of capillary and post
capillary venules .
_ The connective tissue forms a 5th compartment for
neutrophils, but its size is not known.
_ Neutrophils reside for 1-4 days and then die by apoptosis
(whether they have formed their function of phagocytosis
or not )

The appearance of large numbers of immature neutrophils (band cells) in

the blood, sometimes called a “shift to the left,” is clinically significant,
usually indicating a bacterial infection

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 Maturation of lymphocytes & monocytes
_lymphocytes and monocytes are distinguished mainly on the
basis of :
1_ size.
2_ chromatin structure.
3_ the presence of nuclei in smear preparation

As lymphocyte cells mature :

1_ the cells decrease in size.

2_ the chromatin become more compact.
3_ Nucleoli become less visible
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LYMPHOCYTES
_ circulating T lymphocytes originate mainly in the thymus and the
peripheral lymphoid organs ( e.g. spleen , lymph nodes , tonsils )
…..BUT …
All lymphocytes progenitor cells originate in the bone marrow .

_ some of lymphocytes migrate to the thymus, where they acquire

the full attributes of T lymphocytes which populate specific
regions of peripheral lymphoid organs .

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 MONOCYTES
_A large amount of RER is present .

_ An extensive Golgi complex, has a granule condensation

in it which called:

(PRIMARY LYSOSOMES).

_ Mature monocytes enter the blood stream and circulate

for 8 h ………THEN ……..enter the connective tissues
where they mature into MACROPHAGES and function
for several months .

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 Medical application
• Abnormal proliferation of stem cells in bone marrow can produce a range
of myeloproliferative disorders. Leukemias are malignant clones of
leukocyte precursors. They can occur in both lymphoid tissue
(lymphoblastic leukemias) and bone marrow (myelogenous leukemias). In
these diseases, there is usually a release of large numbers of immature
cells into the blood and an overall shift in hemopoiesis, with a lack of some
cell types and excessive production of others. The patient is usually anemic
and prone to infection.

• Diagnosis of leukemias and other bone marrow disturbances involves bone

marrow aspiration. A needle is introduced through the compact bone,
typically at the iliac crest, and a sample of marrow is withdrawn.
Immunocytochemistry with labeled monoclonal antibodies specific to
membrane proteins of precursor blood cells contributes to a more precise
diagnosis of the leukemia.
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Magakaryoblasts

_ it is 15 – 50 µm in diameter .

_ has a large ovoid or kidney shaped nucleus.

_ it has a numerous nuclei, the nucleus becomes highly

polyploid “ contain DNA 30 times more than normal cells
“ before platelets begin to form.

_ the cytoplasm is homogenous and intensely basophilic

_ It has irregularly lobulated nucleus, coarse chromatin and
no visible Nucleoli

_The cytoplasm contains numerous mitochondria, rough

ER, Golgi complex .
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PLATELETS have conspicuous granules originating from Golgi
complex that contains biologically active substances:

1) Platelet derived factor.

2) Fibroblast growth factor.

3) Von Willebrands factor (promotes adhesion of platelets to

endothelial cells )

4) Platelet factor IV ( stimulates blood coagulation)

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Demarcation membranes :

Numerous invaginations of the plasma membranes ramify

through the cytoplasm .

This system defines area of megakaryote’s cytoplasm that

shed platelets, extruding them into the circulation

• Romanowsky-type stain - any stain consisting of methylene

blue and its oxidation products and eosin Y or eosin B.

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