Development of Blood Cells

(Hematopoeisis)

Dr. Zeenat Zaidi

 Composition of Blood
• Blood is classified as a fluid connective tissue
and consists of two main components:
– Plasma, which is a clear extracellular fluid
– Formed elements, which are made up of
the blood cells and platelets
• The formed elements are so named because
they are enclosed in a plasma membrane and
have a definite structure and shape.
• All formed elements are cells except for the
platelets, which are tiny fragments of bone
marrow cells. Include:
– Erythrocytes (red blood cells (RBCs)
– Leukocytes (white blood cells (WBCs). Are
of two types: Granulocytes &
Agranulocytes
– Thrombocytes (Platelets) involved in
clotting
 Hematopoiesis (Hemopoeisis)
• Hematopoiesis is one of the most vital processes in the body, by which all the
blood cellular components are formed.
• It is highly regulated process which:
– Maintains circulating blood cell numbers within relatively constant levels
– Responds to conditions requiring extra cells
– Maintains balance between self renewal, terminal differentiation and cell
death
• Depending on the type of cells, it is further divided into Erythropoeisis
(formation of erythrocytes), Leucopoeisis (formation of leucocytes) and
Thrombopoeisis (formation of thrombocytes).
• In a healthy adult person, approximately 1011–1012 new blood cells are
produced daily in order to maintain steady state levels in the peripheral
circulation

All blood cell types are derived from hematopoieitc stem cells (HSCs)
residing in the bone marrow.
 Hematopoietic Stem Cells (HSCs)
• Hematopoietic stem cells (HSCs) are multipotent cells, residing in the
bone marrow and have the unique ability to differentiate into all mature
blood cell types.
• HSCs are self-renewing. When they proliferate:

Some of their daughter cells

remain as HSCs, so the pool
of stem cells does not
become depleted.

The other daughters cells become the

myeloid and lymphoid progenitor cells,
that can lead to the production of one or
more specific types of blood cells, but
cannot self-renew.
The lymphoid progenitor cells, become lymphoblasts before
differentiating into T cells, B cells, and NK (natural killer) cells.

The common myeloid progenitor cells, differentiate into:

• Myeloblasts
• Erythropoietic proginators
• Magakaryotic progenitors.

The myeloblasts differentiate into:

• Granulocytes
• Monocytes. Monocytes further differentiate
into macrophages upon reaching certain
tissues.
Haematopoiesis
 Hematopoiesis: Locations
• Hematopoiesis begins in early intrauterine life, in the yolk sac
• As development progresses, blood formation occurs in the liver, spleen, and lymph
nodes (extramedullary hematopoeisis).
• The liver becomes the dominant site of hemopoiesis from 3-7 months gestation.
• When bone marrow develops, it eventually assumes the task of forming most of the
blood cells, and by birth, the extramedullary hemopoiesis comes to an end. (If needed,
the liver, thymus, and spleen may resume their hematopoietic function)
• In children, hematopoiesis occurs in the marrow of the long bones, such as the femur
and tibia.
• Through childhood, the red hematopoietic marrow gets invaded by fat cells and become
yellow marrow.
• In adults hematopoietic marrow occurs mainly in the ends of long bones and in the
bones of the axial skeleton i.e., pelvis, cranium, vertebrae, ribs, and sternum (most of
the marrow in long bones is changed to yellow marrow).
• The maturation, activation, and some proliferation of lymphoid cells occurs in secondary
lymphoid organs, such as the spleen, thymus, and lymph nodes.
 Hematopoietic Growth Factors
• Hematopoeisis is tightly controlled by growth factors and the microenvironment.
• Red and white blood cell production is regulated with great precision in healthy humans,
and the production of granulocytes is rapidly increased during infection. The proliferation
and self-renewal of these cells depend on growth factors.
• Growth factors, with the exception of erythropoeitin, are multifunctional. A single growth
factor will influence several different stages of several lineages and promote proliferation,
differentiation, maturation, egress from bone marrow and survival in tissue.
• One of the key players in self-renewal and development of haematopoietic cells is stem
cell factor (SCF). Absence of SCF is lethal.
• Glycoprotein growth factors regulate the proliferation and maturation
• Colony-stimulating factors (CSFs. Three CSFs are granulocyte-macrophage CSF (GM-CSF),
granulocyte CSF (G-CSF) and macrophage CSF (M-CSF). These stimulate granulocyte
formation and are active on either proginator cells or end product cells.
• Erythropoietin, produced in kidney is required for the later stages of erythrocyte
production (has little effect on early stages of RBC production) .
• Thrombopoietin, produced in kidney and liver, makes myeloid progenitor cells
differentiate to megakaryocytes, which produce platelets. It also promotes the early stages
of red cell production.
 Erythropoiesis
• Erythropoiesis is the highly regulated, multistep process by which the body
generates mature red blood cells, or erythrocytes.
• During development, erythropoiesis consists of two major stages:
1. Primitive erythropoiesis, which is initiated in the yolk sac with the
generation of large nucleated erythroblasts and
2. Definitive erythropoiesis, which arises from the fetal liver with the
development of smaller enucleated erythrocytes, that quickly become
dominant in embryonic circulation. The switch of hemoglobin to ‘fetal
types’ also occurs at the initiation of definitive erythropoiesis

• The process of erythropoiesis involves:

– Progressive reduction in size and cytoplasmic organelles
– Inactivation and ultimately extrusion of the nucleus
– Progressive hemoglobin synthesis by ribosomes
 Erythropoiesis cont’d
• The earliest cell in erythropoiesis is the
proerythroblast.
– It is a large cell (15-20µm)
– Shows fine granular nuclear
chromatin containing one or more
paler nucleoli
– High nuclear:cytoplasmic ratio.
– The cytoplasm is often deeply
basophilic due to high amount of RNA
and ribosomes and has a perinuclear
halo. There is No hemoglobin
– Capable of mitosis
• Proerythroblasts divide and develop
into basophilic erythroblasts, or early
normoblasts.
• 14-17µm
• Large nucleus which have coarser
chromatin and absent nucleoli.
• Basophilic cytoplasm
• Hemoglobin synthesis begins. As
hemoglobin production increases,
the cytoplasm develops a grayish
hue.
• Active mitosis
 Erythropoiesis cont’d
• Basophilic erythroblasts develop
into polychromatic erythroblasts
(intermediate normoblasts)
• 10-15µm.
• Have relatively more hemoglobin
in the cytoplasm that stains with
both blue and red dyes
(polychromatia).
• Nucleus has condensed
chromatin in clumps
• No more capable of division.
• Polychromatic erythroblast then
develops into an orthochromatic
erythroblast (late normoblasts)
• (7-10µm)
• The cytoplasm contains abundant
hemoglobin but still contains
ribosomes with continuing synthesis
of hemoglobin, and is stained
predominantly red.
• The nuclear chromatin and nucleus
becomes extremely condensed and
pyknotic.
 Erythropoiesis cont’d

• When the cell is sufficiently

mature, the nucleus is extruded,
and this anuclear red cell is
termed a reticulocyte
• The reticulocyte
– 7.3µm in size
– Has no nucleus
– Has no organelles
– Is larger than the mature RBC
– Is not concave
– Has many polyribosomes
 Erythropoiesis cont’d
• A reticulocyte spends 2-3 days in
the bone marrow before it is
released in the peripheral blood,
where it spends another day to
develop into mature erythrocytes
with loss of the basophilic tinge to
the cytoplasm as RNA levels
decrease.
• Normally 0.5-2% of circulating RBC,
are reticulocytes.
• In severe anemia, many of these
are released into the blood
prematurely (reticulosis)
 Erythropoiesis cont’d

• Incomplete extrusion of nucleus

results in formation of Howell-
Jolly bodies (small, spherical,
condensed nuclear remnants).
• They are normally pinched off in
a small envelope of plasma
membrane by the splenic
macrophages and are not found
in blood.
• After splenectomy, it is common
to find some cells containing
Howell- Jolly bodies in blood
smears
The reticulocyte count is an important diagnostic tool.
• It is a good indicator of bone marrow activity.
• It is used to:
– Differentiate anemias caused by bone marrow failure from those caused by
hemorrhage or hemolysis
– Check the effectiveness of treatment in pernicious anemia and folate and iron
deficiency
– Assess the recovery of bone marrow function in aplastic anemias
– Determine the effect of radioactive substance exposure
• A low reticulocyte count may mean a need for bone marrow biopsy to assess
bone marrow activity
• Reticulocytosis is seen following hemolysis or acute blood loss.
 Erythropoiesis cont’d
Mature Erythrocyte
• Reddish, circular, biconcave cells
• 7-8 µ
• No visible internal structure
• High Hb content
• Bright at centre due to biconcave
shape

Each proerythroblast will generally become 16 RBCs over 5-7 days.

• Differentiation phase: from proerythroblast to reticulocyte phase – 5 days
• Maturation phase: from reticulocyte to RBC – 2 days
• RBC life span= 100 ± 20 days

Abnormal RBCs

Sickle Cell Anemia Spherocytosis

 Regulation of Erythropoiesis
• General factors – Erythropoietin, Growth inducers (IL 1,3,6)
• Maturation factors - Vitamin B 12 - Folic acid
• Factors necessary for hemoglobin production
– Vitamin C: Helps in iron absorption (Fe+++ Fe++)
– Proteins: Amino Acids for globin synthesis
– Iron & copper: Heme synthesis
– calcium, bile salts, cobalt & nickel.
• Stimuli for production:
– Hypoxia
– High altitude
– Anemia
– Chronic lung obstructive diseases
– Catecholamines
– Prostaglandins
– Androgens
 Granulopoeisis
Myeloblast
 Granulopoeisis
• Myeloblast is the earliest
recognizable stage in
granulopoeisis.
• Early myeloblast is a large cell
with a large nucleus containing
open chromatin, several large
nucleoli.
• The basophilic cytoplasm shows
small number of primary
azurophilic (purple) granules.
• Myeloblast gives rise to
promyelocyte
• Promyelocytes are generally larger
than myeloblasts, measuring
approximately 12 to 20 microns.
• The nucleus is similar in size to the
myeloblast The chromatin become
slightly more coarse and clumped
• The nucleoli become less prominent
than in the blast stage
• Cytoplasm is more abundant, with
abundant azurophilic (primary)
granules, some of which cover the
nucleus
• Changes into myelocyte
 Granulopoeisis cont’d
• Myelocytes are cells with a rounded or oval
nucleus that may be flattened at one end.
• Shows progressive condensation of chromatin
and lacks nucleoli.
• Cytoplasm shows development of secondary
or specific granules that are:
– pink in neutrophilic myelocyte
– brick red in eosinophilic myelocyte
– dark blue in basophilic myelocyte.
• The secondary granules increase progressively
with a proportional decrease in primary
granules.
• This process continues through several further
cell division
• Morphological distinction between the three
granulocytic lines is first evident at the
myelocyte stage.
 Granulopoeisis cont’d
• Metamyelocytes:
• It is an end cell, incapable of cell division.
• It begins nuclear segmentation with
increasingly indented nucleus.
• The nucleus becomes horse-shoe shaped and
the cell is called ‘stab cell’ or ‘band forms’.
• These immature neutrophils enter the
functional reserve pool in the marrow.
• On entry to the circulation:
– Only half of these band form circulate
– The other half adhere to the
endothelial walls of the small vessels,
forming a marginal pool. These pools
provide reserve sources and are
mobilized on demand
• In the circulation, neutrophils show
progressive segmentation of their nucleus
into 2 lobes (young cells), 3-4 lobes (average
cells) and 5 lobes (older cells)
 Granulopoeisis cont’d
• Neutrophils are highly motile
cells, leave the circulation in
response to chemotactic signals
generated by inflammation. They
do not enter the blood stream
from tissue, but undergo lysis in
tissues
Duration of Granulopeisis:
• The normal developmental
process in marrow from the
myeloblast to myelocyte takes 6
days, and from mylocyte to
release of neutrophils in the
blood another 7 days.
• Production is driven by a range of
growth factors and cytokines
 Barr Body

• Some neutrophils in
females show a small,
dense, dark-staining
drumstick-like projection on
one of the its lobes. This is
called a Barr body
• Barr body is an inactivated,
condensed X chromosome
and is regarded as
diagnostic of genetic
femaleness.
 Monopoeisis
• Monoblasts are normally found in bone
marrow and do not appear in the
normal peripheral blood.
• They are large cells, with round to oval
nuclei and abundant basophilic
agranular cytoplasm.
• The chromatin pattern is very fine, and
nucleoli are prominent.
• Promonocyte: cytoplasmic granules are
developed and the cytoplasm gives a
frosty-glass appearance. Nucleolie less
prominent.
• The promonocytes exit the bone
marrow and travel to lymphoid tissue
where they differentiate into
monocytes. Some monocytes develop
into macrophages.
• A typical promonocyte will undergo two
serial cell division to produce 4
monocytes in a process taking about 60
hours.
 Lymphopoeisis
• Lymphocyte precursors, lymphoblasts
originate in the bone marrow.
• Lymphoblasts are large cells, with fine
open nuclear chromatin, a few pale
nucleoli and scant cytoplasm. These cells
typically differentiate to form
prolymphocytes and mature
lymphocytes.
• B cells and natural killer (NK) cells
complete most of their development
within the bone marrow
• T cells are generated in the thymus from
precursor cells that migrate from the
bone marrow.
• Normally lymphoblasts are found in the
bone marrow, but in acute lymphoblastic
leukemia (ALL), lymphoblasts proliferate
uncontrollably and are found in large
numbers in the peripheral blood.
 Thrombopoiesis (Platelet Formation)
• Platelets are continuously produced as a
component product of hematopoiesis (blood
cell formation).
• Thrombopoiesis occurs from common
myeloid progenitor cells in the bone marrow,
which differentiate into promegakaryocytes
and then into megakaryocytes.
• Megakaryocytes stay in the bone marrow
and, when mature, extrude pseudopodia
known as protoplatelets, into the bone
marrow sinusoids
• The protoplatelets then break up into
hundreds of platelets that circulate
throughout the bloodstream, while the
remaining nucleus of the ruptured
megakaryocyte is consumed by macrophages.
 Thrombopoiesis (cont’d)

• Megakaryocyte and platelet production is

regulated by thrombopoietin.
• Each megakaryocyte produces between
5,000 and 10,000 platelets before its
cellular components are fully depleted.
• Altogether, around 1011 platelets are
produced each day in a healthy adult.
• The average lifespan of a platelet is just 5
to 10 days.
• Old platelets are destroyed by macrophage
phagocytosis in the spleen and by Kupffer
cells in the liver.
• Up to 40% of platelets are stored in the
spleen as a reserve, released when needed
by sympathetically-induced splenic muscle
contractions during severe injury.
 Implications of Platelet Formation
• Balanced thrombopoiesis is important because it directly influences the amount of
platelets in the body and their associated complications.
• An abnormality or disease of the platelets is called a thrombocytopathy, which could be
either a:
• Low number of platelets (thrombocytopenia): If the number of platelets is too low,
excessive bleeding can occur, even from minor or superficial injuries.
– Decrease in function of platelets (thrombasthenia)
– Increase in the number of platelets (thrombocytosis). If the number of platelets is
too high, blood clots can form (thrombosis) and travel through the bloodstream,
which may obstruct blood vessels and result in ischemic events (stroke, myocardial
infarction, pulmonary embolism, or infarction of other tissues).
• Variations with the number of circulating platelets is often due to:
– Issues in thrombopoietin feedback regulation
– genetic characteristics
– Certain medications
– Diseases e.g. thrombocytopenia often occurs in leukemia patients.
 Leukemia

• A cancer found in the blood and bone marrow, caused by abnormal rapid
formation of white blood cells in the body. The excessive WBCs don’t let
the body fight disease and prevent the body from making RBCs and
platelets, resulting in bleeding and anemia.