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.