Hematopoiesis
OVERALL BLOOD CELL MATURATION & FUNCTION
HEMATOPOIESIS - blood cell production
5 y/o - the bone marrow is red & cellular & actively producing
cells
5 - 7 y/o - inactive long bones & fat cells appear to replace the
active marrow (red marrow is gradually transforming into yellow
marrow)
18 - 20 y/o - red marrow remains only in the vertebrae, ribs,
sternum, skull & partially in the femur & humerus
Hematopoiesis
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.
STAGES OF HEMATOPOIESIS
PRENATAL HEMATOPOIESIS
ADULT HEMATOPOIESIS
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
PE – produces Hb
- Gower I
Gower II three embryonic forms of Hb
Portland
PRENATAL HEMATOPOIESIS
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:
erythropoiesis- until the end of normal gestation (splenic)
myelopoiesis – but becomes minimal by the 5th month
Lymphopoiesis – lifetime
d. Lymph nodes
- NRBC’s , Granulocytes, Monocytes, Lymphocytes and Megakaryocytes
Hb A
Hb A2 hemoglobins
Hb F
PRENATAL HEMATOPOIESIS
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
ADULT HEMATOPOIESIS
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, scapulae, sternum, skull
and proximal extremities of the long bones
The Bone Marrow
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
The Bone Marrow
RED MARROW – 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.
Red Bone Marrow
Site of Blood
Yellow Bone Marrow Cell Formation
Red Bone Marrow
Graphic illustration of the arrangement of the extravascular area in hematopoietic
tissue
Adipocytes
Hematopoietic
Tissue
Bone
The Bone Marrow
Normal Bone Marrow
Full of developing
precursor cells in all
stages of maturation
Released into blood at
maturity
ONLY mature cells are
released normally
First recognizable
precursor in each cell
line is a blast
The Bone Marrow
Marrow recedes
during development
of the individual,
and in the adult
occupies only the
support skeleton and
the proximal regions
of the long bones
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
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
Spleen
Reticuloendothelial organ
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
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
Thymus
Thymus Gland
responsible for normal development of some
of the lymphocytes
located in the neck
maximum development in childhood, atrophies
with age
Bursa Fabricus
foundin birds with possible analogous tissue in
man. Responsible for normal antibody
production
Thymus
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
Thymus
CFU-T
T
Lymphocytes
CFU-L B
CFU-B
Bursa or Bone Marrow
Nuetrophils
CFU-
GM
THSC Monocytes
Macrophages
Eosinophils
CFU-
EO
Basophils
CFU-S
CFU-
MEG
Platelets
BFU-E CFU-E Erythrocytes
Kierszenbaum, Fig. 6-16; See
Hematopoiesis Ross 5th Ed Table 10.4
PPSC
Hematopoiesis
Regulation of Hematopoiesis
Erythropoietin (EPO) - major regulator of erythropoiesis, stimulates
erythroid CFU cells and proerythroblasts
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
TERMINOLOGY
Pluripotent hematopoietic stem cell (HSC)
Undifferentiated cell producing blood cells of all lineages,
capable of self-renewal (embryonal stem cell)
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)
HEMATOPOIESIS
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
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
ERYTHROPOIESIS:
Erythrocyte Normal Maturation Series
Pronormoblast
(Rubriblast)
(Proerythroblast)
Size: 12-25 um in diameter
Cytoplasm:
Deeply basophilic
Relatively small amount
Perinuclear halo
Nucleus
Large
Round to sl. Oval
Reddish purple
1-3 nucleoli
N/C ratio: 8:1
1% of nucleated cells in bone marrow
Erythrocyte Normal Maturation Series
Basophilic Normoblast
(Prorubricyte)
(Basophilic Erythroblast)
Size: 12-17 um in diameter
Cytoplasm: intensely basophilic
Nucleus:
Relatively large
Round to slightly oval
Coarser chromatin
Indistinct nucleoli
Occupies 75% of the cell
1-3% of nucleated cells of bone
marrow
Erythrocyte Normal Maturation Series
Polychromatophilic Normoblast
(Rubricyte)
(Polychromatophilic Erythroblast)
Size: 12-15 um in diameter
Cytoplasm: blue-gray to pink gray
(production of hemoglobin)
Nucleus:
Round, eccentric
Smaller
More condensed
Stains deeper blue-purple
13-30% of nucleated cells in
bone marrow
This is the last cell division during
maturation
Erythrocyte Normal Maturation Series
Orthochromic Normoblast
(Metarubricyte)
(Orthochromic Erythroblast)
Size: 8-12 um in diameter
Cytoplasm: pinker, increased amount
of Hb
Nucleus:
Pyknotic
eccentric
1-4% of nucleated cell in bone
marrow
Nucleus is extruded at this stage
Erythrocyte Normal Maturation Series
Reticulocyte
Diffusely Basophilic Erythrocyte*
Polychromatophilic Erythrocyte*
Size: 7-10 mm in diameter
Cytoplasm:
Pink to pinkish grey
Still contains small amounts
of RNA (polychromasia)
Nucleus: none
Within 24-48 hrs, the cell loses
the organelles & assumes a
biconcave shape
Erythrocyte Normal Maturation Series
Erythrocyte
Size: approximately 7.2 um in diameter
Cytoplasm: Pink
The red blood cell is non-nucleated,
round and biconcave
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.
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:
Erythropoiesis
KIDNEY LIVER BONE
MARROW
Store Iron, Site of RBC’s
Erythropoietin Store protein, Vit. B12 & Folic, formation
Synthesize globin,
Produce eryhtropoietin (10%)
Regulation of Erythropoiesis
Too few RBCs leads to tissue hypoxia
Too many RBCs increases blood
viscosity
Balance between RBC production and
destruction depends on:
Hormonal controls
Adequate supplies of iron, amino
acids, and B vitamins
LEUKOPOIESIS: PRODUCTION OF LEUKOCYTES
• Leukopoiesis is 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
• Many hematopoietic hormones are used
clinically to stimulate bone marrow
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
Band Stage
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.
Granulopoiesis – Development of Granulocytes
46 Earliest recognizable cell in marrow, large euchromatic
nucleus, nucleoli, basophilic cytoplasm
Large spherical nucleus, azurophilic or primary cytoplasmic
granules (only produced by these cells), no subtypes ( can't
recognize cell lines at this stage)
Spherical nucleus, heterochromatic, specific or secondary
granules (seen only in granulocytes) begin to form, can
divide
Easily recognizable lineages, kidney bean –shaped
nucleus
Not seen in Eosinophil or Basophil lineage, also called
stab form, horse-shoe shape nucleus, up to 3% in
peripheral blood
Mature forms
LYMPHOPOIESIS
Lymphocytes are derived from committed
stem cells that originate from pluripotent
stem cell.
Early lymphoid cells further differentiates
into B & T lymphocytes.
LYMPHOCYTIC MATURATION SEQUENCE
LYMPHOBLAST
Cell is similar to other blast cells. It is round or oval, very large, with a large round to oval reddish-
purple nucleus.
The nuclear chromatin material is fine and well distributed but perhaps more coarse than in
myeloblasts.
The nucleus contains one or two nucleoli.
The cytoplasm is bluish and non granular and forms a thin rim around the nucleus.
PROLYMPHOCYTE
The nucleus is round or oval in shape but smaller than the lymphoblast.
The nuclear chromatin is coarse and slightly clumped.
Nucleoli or remnants of nucleoli may be present.
There is an abundant amount of light blue cytoplasm around the nucleus. Also, there may be a few
azurophilic granules in the cytoplasm.
LYMPHOCYTE
This is the mature cell of the lymphocytic series and the only cell form found in the peripheral blood
Lymphocytes vary greatly in size and may be classified as small, medium or large. However, size does not
determine age of these cells.
The cells are easily distorted and often appear in irregular shapes in stained preparations. The nuclear
chromatin is condensed to form large, discrete almost solid clumps, with thickening of the nuclear
membrane. Nucleoli are absent. Non specific granules may be observed in the cytoplasm of these cells.
LYMPHOPOIESIS
Immunologically competent cells
Primary lymphoid organs
Bonemarrow
Thymus
Secondary lymphoid organs
Lymph nodes
Spleen
Lymphoid tissues
Lymphocytes
B and T lymphocytes
NK killer cells
MONOPOIESIS
Development of the monocyte
Stages in the monocytic development
are:
Monoblast
Promonocyte
Monocyte
WHITE BLOOD CELL
Neutrophil
- 2-5 lobe nucleus
- Primary or secondary
granules
Pink (azurophilic granules)
Grey-blue granules
- Life 10 hours
Precursors:
Myeloblast <4%
Pro myelocytes
Myelocytes
Metamyelocytes
Band form (stab form)
WHITE BLOOD CELL
•Eosinophil
- Coarser & more
deeply red staining
granules
- Rarely more than
two lobes of nucleus
- Special role in
allergy, inflamation
& parasite infection
WHITE BLOOD CELL
•Basophil
- Occasionally seen
- Dark cytoplasmic
granules
- Role in
hypersensitivity
response
- Give rise to mast
cells
WHITE BLOOD CELL
Monocyte Larger than lymphocyte
Oval or indented nucleus
Monocytes >>>>to
macrophage
Specific function depends on
the tissue type:
Kupffer, Oestoclast, Microglia,
Serosal, Langerhans
WHITE BLOOD CELL
Lymphocyte
• Nucleus is dense, round, oval
or slightly indented
• B lymphocyte - humoral
immunity (~20-30%)
• T lymphocyte - cell-mediated
immunity (~60-80%)
• Natural killer (NK) cell - cell-
mediated immunity (~5-10%)
• Agranulocyte - lysosomal acid
hydrolases
4,000-11,000 cells/mm 3
Granulocytes
Neutrophils- 40-70%
Eosinophils- 1-4%
Basophils- <1%
Agranulocytes
Monocytes- 4-8%
Lymphocytes- 20-45%
Leukocytes
Leukocyte
Mast
cell
Distribution - blood and CT (as transient or wandering cells)
Function - immune protection, movement (cell motility)
Diapedesis - movement out of blood into connective tissue
Chemotaxis - movement directed by homing molecules
ID WBC’s
eosinophil
neutrophil
monocyte
RBC
neutrophil
monocyte
lymphocyte
lymphocyte
basophil
THROMBOPOIESIS: GENESIS OF PLATELETS
• The stem cell for platelets is the hemocytoblast
• The sequential developmental pathway is as
shown:
Stem Cell Developmental pathway
Hemocytoblast Megakaryoblast Promegakaryocyte Megakaryocyte Platelets
Stages in Thrombocyte Development
MEGAKARYOBLAST
The cell is large, irregularly shaped with a single or
several round or oval nuclei and with a blue, non
granular cytoplasm. Nucleoli are usually present.
Stages in Thrombocyte Development
PROMEGAKARYOCYTE
This cell differs from the
megakaryoblast in that there
are bluish granules in the
cytoplasm adjacent to the
nucleus. The nucleus in this
second stage of maturation
has usually divided one or
more times and the cell has
increased in size.
Stages in Thrombocyte Development
MEGAKARYOCYTE
The cell is very large with
relatively large amounts of
cytoplasm, and multiple
nuclei. The cytoplasm contains
numerous small, uniformly
distributed granules that are
reddish-blue in color.
