Hematopoiesis

Introduction

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. Most bone marrow cells are morphologically recognizable
precursors of granulocytes or erythrocytes with smaller numbers of platelet precursors
(megakaryocytes), lymphocytes, monocytes, macrophages, stromal cells (endothelial cells, fibroblasts,
osteoblasts, and osteoclasts), eosinophils, plasma cells, basophils, mast cells, and blasts. The latter
include hematopoietic stem cells (HSCs), which are capable of self-renewal and differentiation; and
progenitors, which differentiate along a specific pathway.

Blood cell production (hematopoiesis) encompasses the overall interactions of cellular

proliferation, differentiation, morphogenesis, functional maturation and death. Basically, the
development pattern of embryogenesis that involves cellular differentiation is an orderly process of
many morphologic and functional changes for which the regulatory mechanism is not understood. Many
of the developmental patterns occur simultaneously in the embryo and fetus.

Red bone marrow is a highly vascularized connective tissue located in the microscopic spaces
between trabeculae of spongy bone tissue. It is present chiefly in bones of the axial skeleton, pectoral an
pelvic girdles, and the proximal epiphyses of the humerus and femur. About 0.05–0.1% of red bone
marrow cells are derived from mesenchyme and are called pluripotent stem cells (ploo-RIP-o¯ -tent;
pluri- _ several) or hemocytoblasts. These cells have the capacity to develop into many different types of
cells. In newborns all bone marrow is red and thus active in blood cell production. As an individual grows
and in adulthood, the rate of blood cell formation decreases; the red bone marrow in the medullary
(marrow) cavity of long bones becomes inactive and is replaced by yellow bone marrow, which is largely
fat cells. Under certain conditions, such as severe bleeding, yellow bone marrow can revert to red bone
marrow by extension of red bone marrow into yellow bone marrow and repopulation of yellow bone
marrow by pluripotent stem cells.

Stem cells in red bone marrow reproduce themselves, proliferate, and differentiate into cells
that give rise to blood cells, macrophages, reticular cells, mast cells, and adipocytes. Some of the stem
cells can also form osteoblasts, chondroblasts, and muscle cells, and someday may be used as a source
of bone, cartilage, and muscular tissue for tissue and organ replacement. The reticular cells produce
reticular fibers, which form the stroma (framework) that supports red bone marrow cells. Once blood
cells are produced in red bone marrow, they enter the bloodstream through sinusoids (also called
sinuses), enlarged and leaky capillaries that surround red bone marrow cells and fibers. With the
exception of lymphocytes, formed elements do not divide once they leave red bone marrow.

In order to form blood cells, pluripotent stem cells in red bone marrow produce two further
types of stem cells, which have the capacity to develop into several types of cells. These stem cells are
called myeloid stem cells and lymphoid stem cells. Myeloid stem cells begin their development in red
bone marrow and give rise to red blood cells, platelets, monocytes, neutrophils, eosinophils, and
basophils. Lymphoid stem cells begin their development in red bone marrow but complete it in
lymphatic tissues; they give rise to lymphocytes. Although the various stem cells have distinctive cell
identity markers in their plasma membranes, they cannot be distinguished histologically and resemble
lymphocytes.

During hemopoiesis, some of the myeloid stem cells differentiate into progenitor cells (pro¯-
JEN-i-tor). Other myeloid stem cells and the lymphoid stem cells develop directly into precursor cells
(described shortly). Progenitor cells are no longer capable of reproducing themselves and are committed
to giving rise to more specific elements of blood. Some progenitor cells are known as colony-forming
units (CFUs). Following the CFU designation is an abbreviation that indicates the mature elements in
blood that they will produce: CFU–E ultimately produces erythrocytes (red blood cells), CFU–Meg
produces megakaryocytes, the source of platelets, and CFU–GM ultimately produces granulocytes
(specifically, neutrophils) and monocytes. Progenitor cells, like stem cells, resemble lymphocytes and
cannot be distinguished by their microscopic appearance alone.

In the next generation, the cells are called precursor cells, also known as blasts. Over several
cell divisions they develop into the actual formed elements of blood. For example, monoblasts develop
into monocytes, eosinophilic myeloblasts develop into eosinophils, and so on. Precursor cells have
recognizable microscopic appearances.

Several hormones called hemopoietic growth factors regulate the differentiation and
proliferation of particular progenitor cells. Erythropoietin (e-rith_-ro¯-POY-e-tin) or EPO increases the
number of red blood cell precursors. EPO is produced primarily cells in the kidneys that lie between the
kidney tubules (peritubular interstitial cells). With renal failure, EPO release slows and RBC production is
inadequate. Thrombopoietin (throm_-bo¯-POY-e¯ -tin) or TPO is a hormone produced by the liver that
stimulates the formation of platelets (thrombocytes) from megakaryocytes. Several different cytokines
regulate development of different blood cell types. Cytokines are small glycoproteins that are typically
produced by cells such as red bone marrow cells, leukocytes, macrophages, fibroblasts, and endothelial
cells. They generally act as local hormones (autocrines or paracrines; see Chapter 18). Cytokines
stimulate proliferation of progenitor cells in red bone marrow and regulate the activities of cells
involved in nonspecific defenses (such as phagocytes) and immune responses (such as B cells and T
cells). Two important families of cytokines that stimulate white blood cell formation are colony-
stimulating factors (CSFs) and interleukins.

Hemopoietic growth factors made available through recombinant DNA technology hold
tremendous potential for medical uses when a person’s natural ability to form new blood cells is
diminished or defective. The artificial form of erythropoietin (Epoetin alfa) is very effective in treating
the diminished red blood cell production that accompanies end-stage kidney disease. Granulocyte-
macrophage colony-stimulating factor and granulocyte CSF are given to stimulate white blood cell
formation in cancer patients who are undergoing chemotherapy, which kills red bone marrow cells as
well as cancer cells because both cell types are undergoing mitosis. (Recall that white blood cells help
protect against disease.) Thrombopoietin shows great promise for preventing the depletion of platelets,
which are needed to help blood clot, during chemotherapy. CSFs and thrombopoietin also improve the
outcome of patients who receive bone marrow transplants. Hemopoietic growth factors are also used to
treat thrombocytopenia in neonates, other clotting disorders, and various types of anemia. Research on
these medications is ongoing and shows a great deal of promise.
3 Developmental Periods
Mesoblastic Period

Hematopoiesis begins during embryonic development in the blood islands of the yolk sac. These
blood islands are first detected at approximately 19-20 days of gestation, and their appearance marks
the beginning of the mesoblastic period of hematopoiesis. Most likely, blood islands develop from the
mesodermal embryonic layer of the yolk sac. The blood islands remain active only through the 18 to 12
week of gestation and are primarily responsible for red cell production (erythropoiesis), although areas
of leukopoiesis and platelet production (megakaryopoiesis) have been identified.

Hepatic Period

Definitive morphologic hematopoiesis begins during the fifth to sixth weeks of gestation in the
liver and marks the beginning of the hepatic period of hematopoiesis. At this point cells appear that are
morphologically identifiable, the liver remains the primary site of blood cell production until the sixth
month and may continue to produce blood cells, although to a much smaller degree, until the first or
second week after birth. Because liver hematopoiesis is intravascular, infants normally have a few
circulating nucleated red cells. At first the fetal liver is primarily an erythroid organ and produces red
cells containing fetal hemoglobin. The production of granulocytes and lymphocytes is minimal in the
fetal liver. During the hepatic period, the spleen, thymus, and lymph nodes also become active in blood
cell production.

Myeloid period

During the fifth month of gestation, bone cavities begin to form, and bone marrow begins to
assume responsibility as the main site of blood cell production. This marks the beginning of the myeloid
period. During this time hemoglobin A1 consisting of two alpha and two beta globin chains, begins to
appear and gradually increase in concentration. Most adult hemoglobin is Hb A1.

After the first three weeks postpartum, the bone marrow become the only normal site of blood
cell production and remains so throughout life. During the first few years of life, a delicate balance exists
between developing bone marrow space and developing infant’s need for blood cells.

Erythropoiesis: Production of RBCs

Erythropoiesis (e-rith_-ro¯-poy-EE¯ -sis), the production of RBCs, starts in the red bone marrow
with a precursor cell called a proerythroblast. The proerythroblast divides several times, producing cells
that begin to synthesize hemoglobin. Ultimately, a cell near the end of the development sequence ejects
its nucleus and becomes a reticulocyte (re-TIKu ¯-lo¯-sı¯t). Loss of the nucleus causes the center of the
cell to indent, producing the red blood cell’s distinctive biconcave shape. Reticulocytes retain some
mitochondria, ribosomes, and endoplasmic reticulum. They pass from red bone marrow into the
bloodstream by squeezing between the endothelial cells of blood capillaries. Reticulocytes develop into
mature red blood cells within 1 to 2 days after their release from red bone marrow.

Normally, erythropoiesis and red blood cell destruction proceed at roughly the same pace. If the
oxygen-carrying capacity of the blood falls because erythropoiesis is not keeping up with RBC
destruction, a negative feedback system steps up RBC production. The controlled condition is the
amount of oxygen delivered to body tissues. Cellular oxygen deficiency, called hypoxia (hı¯-POKS-e¯ -a),
may occur if too little oxygen enters the blood. For example, the lower oxygen content of air at high
altitudes reduces the amount of oxygen in the blood. Oxygen delivery may also fall due to anemia, which
has many causes: Lack of iron, lack of certain amino acids, and lack of vitamin B12 are but a few.
Circulatory problems that reduce blood flow to tissues may also reduce oxygen delivery. Whatever the
cause, hypoxia stimulates the kidneys to step up the release of erythropoietin, which speeds the
development of proerythroblasts into reticulocytes in the red bone marrow. As the number of
circulating RBCs increases, more oxygen can be delivered to body tissues.

Premature newborns often exhibit anemia, due in part to inadequate production of

erythropoietin. During the first weeks after birth, the liver, not the kidneys, produces most EPO. Because
the liver is less sensitive than the kidneys to hypoxia, newborns have a smaller EPO response to anemia
than do adults. Because fetal hemoglobin (hemoglobin present at birth) carries up to 30% more oxygen,
the loss of fetal hemoglobin, due to insufficient erythropoietin production, makes the anemia worse.

Leukopoiesis

Leukopoiesis, the process of making leukocytes, is stimulated by various colony-stimulating

factors (CSFs), hormones produced by mature white blood cells. The development of each kind of white
blood cell begins with the division of hemopoietic stem cells into one of the following “blast”cells.

● Myeoblasts divide to form eosinophilic, neutrophilic, or basophilic myelocytes, which lead to the
development of the three kinds of granulocytes.
● Monoblasts lead to the development of monocytes.
● Lymphoblasts lead to the development of lymphocytes.

Myeloblast Promyelocyte Myelocyte

 Size 15-20 um 15-20 um 12-18 um

Moderate in amount

Increase in amount Few non-specific

Small in amount
granules
Pale blue
Cytoplasm Moderate blue in color
Small amount of
With few non-
Smooth & granular specific granules
specific granules
begin to appear
(N,E,B)

Oval or round
Round or sl oval Oval or round
CP coarser and more
Fine CP CP coarser & more
clumped
Reddish purple clumped
Nucleus Centrally located
4/5 of the cell Centrally located
½ of the cell
N/C ratio: 4:1 N/C ratio: 1:1
N/C ratio: 3:1
With 2-5 nucleoli No nucleolus
2-3 nucleoli

Metamyelocyte Band Cell Segmenters

Size 10-15 um 9-15 um 12-18 um

Moderate to abundant
N-pink to rose-purple
amount
granules
Few non-specific Same as
Cytoplasm E-reddish orange
granules metamyelocyte
B-dark purple to blue
Full specific granules
black granules
(N,E,B)

Nucleus Indented or kidney- Elongated or band- N: 2-5 lobes connected

shaped shaped by thin nuclear
filament
CP is coarse & clumped Deeply indented
 E: 2 lobes
CP is coarse &
Stains dark blue B: unsegmented or 2
clumped
lobes

Lymphocytic Series

Lymphoblasst Prolymphocyte Mature Lymphocyte

Small 8-10 um

Size 10-18 um 10-18 um Medium 10-12 um

Large 12-16 um

Small: thin rim around

the nucleus (dark
Smooth, no granules blue)
Usually non-granular,
Moderate to dark blue but with few Medium: more
Cytoplasm
(dark to lighter near occasional abundant (moderate
the nucleus) azurophilic granules blue)

Large: abundant (very

pale blue)

S: CP is clumped, sl
indented
CP is coarse CP is more clumped
and coarser M: CP is dense, sl
Round or oval indented
Nucleus Round or slightly
1-2 nucleoli indented L: CP is coarse, sl
indented,
N/C ratio: 4:1 1-2 nucleoli eccentrically located

No nucleoli

Monocytic Series
 Monoblast Promonocyte Monocyte

Size 12-20 um 14-18 um 12-20 um

Blue-gray
Blue-gray
Many azurophilic
Contains few fine granules
Moderately basophilic
azurophilic ganules
Cytoplasm to blue-gray Ground glass
Ground glass appearance
Non-granular
appearance
Vacuoles present
Moderate in amount
Abundant in amount

Ovoid or round in Round, kidney-shaped

shape Oval or may have 1 with lobulations
single fold folded over the top,
Light blue-purple
showing brain-like
Nucleus With fine, lacey Fine CP convolutions
chromatin 1-2 nucleoli CP not clumped, lacey
1-2 nucleoli and fine
N/C ratio: 3:1
N/C ratio: 4:1 No nucleoli
Thrombopoiesis
Thrombopoiesis, the process of making platelets, begins with the formation of megakaryoblasts
from hemopoietic stem cells. The megakaryoblasts divide without cytokinesis to become
megakaryocytes, huge cells with a large, multilobed nucleus. The megakaryocytes then fragment into
segments as the plasma membrane infolds into the cytoplasm.
 Granular
Megakaryoblast Promegakaryocyte
Megakaryocyte

Size 20-50 um 20-60 um 30-90 um

Varying shade of blue

Pinkish blue
W/ small pseudopods Less basophilic
Abundant
Cytoplasm Small in amount (narrow Abundant
With irregular border
band) Granules start to form
With fine granules
Non-granular

Round, oval or kidney Irregular in shape with

Multilobulated
shaped sl lobulation
CP is coarser
Nucleus Fine CP CP is coarse
No nuceloli
Multiple nucleoli Multiple nucleoli
N/C ratio: 2:1
N/C ratio: 10:1 N/C ratio: 4:1 to 7:1

Mature Megakaryocyte Platelets or Thrombocytes

Size 40-120 um 1-4 um

Light to purple
With coarse clumps of
Very granular
Cytoplasm granules aggregating into small
bundles, which bud off 2 parts: chromomere,
hylamore

Multiloblated

Nucleus No nucleoli none

N/C ratio: 1:1

Questions for Research:
1. Describe changes that takes place in the following as the cell matures:
1.1 Cell size
⮚ As the cell matures, the cell size decreases
1.2 Nucleus (erythropoiesis)
⮚ In erythropoiesis, as the cell matures the nucleus becomes smaller and finally
extruded.
1.3 Nucleus (leukopoiesis)
⮚ In leukopoiesis, as the cell matures the nucleus from large to small and then
undergoes segmentation and lobulation
1.4 Amount of cytoplasm
⮚ The amount of cytoplasm becomes larger due to the change in size of the nucleus
from large to small.
1.5 Basophilia of cytoplasm
⮚ In the case of the RBC, the cytoplasm becomes less basophilic due to RNA
degeneration and production of hemoglobin.
1.6 Staining characteristics of the cytoplasm
⮚ In the early stages of cell maturation, the cytoplasm of the cell is nonspecific to any
stain but as the cell matures, it starts of develop affinity to a particular stain.
1.7 Chromatin pattern
⮚ As the cell matures, the chromatin pattern changes from fine & delicate to course
and clumped.

2. Enumerate sites where adult hematopoiesis may take place.

⮚ At the age of 18 years old, the only active hematopoietic sites are in the pelvis,
vertebrae, ribs, sternum, skull and proximal extremities of the long bone.
 3. Newborn babies would have higher blood cell count than adults. Why?
⮚ In newborns all bone marrow is red and thus active in blood cell production. As an
individual grows and in adulthood, the rate of blood cell formation decreases; the
red bone marrow in the medullary (marrow) cavity of long bones becomes inactive
and is replaced by yellow bone marrow, which is largely fat cells. Under certain
conditions, such as severe bleeding, yellow bone marrow can revert to red bone
marrow by extension of red bone marrow into yellow bone marrow and
repopulation of yellow bone marrow by pluripotent stem cells.

4. Enumerate 3 stages of intrauterine hematopoiesis and identify the organ involved in blood cell
formation.
⮚ Mesoblastic stage – Blood islands of the yolk sac
⮚ Hepatic stage – liver is the main erythroid organ
⮚ Myeloid stage - bone marrow is the site of blood cell production

5. Differentiate extramedullary and intramedullary hematopioesis.

⮚ Extramedullary hematopoiesis is the production of blood cells from the different organs
aside from the bone marrow. Examples of which are the lymph nodes and the spleen.
While Intramedullary hematopoiesis is the production of blood cells from the bone
marrow alone.

6. Identify one condition that may be associated with an increase of:

6.1 Erythropoiesis – Polycythemia vera
6.2 Leukopoiesis - Leukemia
6.3 Thrombopoiesis - Thrombosis

References:
Lotspeich-Steininger, Cheryl. Clinical Hematology Principles, Procedures, Correlations
Henry, John Bernard. Clinical Diagnosis and Management by Laboratory Methods 22 nd edition
Tortora, Gerard. Principles of Anatomy & Physiology 12 th edition
G. Moore et. Al, Haematology, 2010
http://www.scribd.com/doc/50704881/110/Leukopoiesis