BY:

PAULAN, MARY ABIGAIL, RMT

HEMATOPOIESIS
▪ A continuous, regulated process of blood cell production
that include cell renewal, proliferation, differentiation
and maturation.
Mesoblastic phase
▪ YOLK SAC
▪ Site of primitive hematopoiesis
▪ blood islands
▪ contain predominantly primitive erythroblasts
▪ formed during the first 2 to 8 weeks of life.
▪ produce hemoglobin (Gower-1, Gower-2, and Portland)
Hepatic Phase
▪ LIVER
▪ sixth week ( 2nd – 5th month)
▪ major hematopoietic organ of early and midfetal life
▪ gives rise to fetal Hgb
▪ In the middle part of fetal life, the spleen and, to a
lesser extent, lymph nodes have a minor role in
hematopoiesis
 Medullary phase
▪ Bone marrow
▪ Begins in the 4th month of gestation
▪ After the fifth fetal month  the primary site of hematopoiesis
▪ Until post-natal life
BONE MARROW
▪ yellow marrow  normally inactive, composed mostly of fat
(adipose) tissue
▪ red marrow  normally active in the production of most types
of leukocytes, erythrocytes, and thrombocytes
▪ At birth  dominated by red marrow
▪ By age 18, red marrow is found only in the vertebrae, ribs,
sternum, skull bones, pelvis, and to some extent the proximal
epiphyses of the femur and humerus
HEMATOPOIETIC
MICROENVIRONMENT
1. Endothelial cells
2. Adipocytes
3. Macrophages
4. Osteoblasts
5. Osteoclasts
6. fibroblasts
 HEMATOPOIETIC
MICROENVIRONMENT
▪ Endothelial cells
▪ broad, flat cells that form a single continuous layer along the inner
surface of the arteries, veins, and vascular sinuses.
▪ regulate the flow of particles entering and leaving hematopoietic spaces in
the vascular sinuses
▪ Adipocytes
▪ are large cells with a single fat vacuole
▪ they play a role in regulating the volume of the marrow in which active
hematopoiesis occurs
 HEMATOPOIETIC MICROENVIRONMENT
▪ Macrophages
▪ function in phagocytosis, and both macrophages and lymphocytes
secrete various cytokines that regulate hematopoiesis
▪ they are located throughout the marrow space
▪ Osteoblasts
▪ bone-forming cells
▪ Osteoclasts
▪ bone-resorbing cells.
▪ Reticular adventitial cells
▪ form an incomplete layer of cells on the abluminal surface of the
vascular sinuses
EXTRAMEDULLARY
HEMATOPOIESIS
▪ abnormal circumstances, the spleen, liver, and lymph nodes revert
back to producing immature blood cells
1. When the bone marrow becomes dysfunctional in cases such
as aplastic anemia, infiltration by malignant cells, or over
proliferation of a cell line (e.g., leukemia).
2. When the bone marrow is unable to meet the demands
placed on it, as in the hemolytic anemia
PROGENITOR BLOOD CELLS
▪ multipotential hematopoietic stem cell
▪ Progenitor of all blood cells
 LINEAGE SPECIFIC HEMATOPOIESIS
▪ Erythropoiesis occurs in distinct anatomical sites called erythropoietic
islands, specialized niches in which erythroid precursors proliferate,
differentiate, and enucleate.
▪ Granulopoiesis can be recognized as a maturational unit. Maturing
cells spend an average of 3 to 6 days in the proliferating pool. average life
span of 6 to 10 hours in circulation
▪ lymphocytes and plasma cells are produced in lymphoid follicles
▪ Megakaryopoiesis takes place adjacent to the sinus endothelium.
develop into platelets in approximately 5 days.
INTERLEUKINS
▪ cytokines that act independently or in conjunction with
other interleukins to encourage hematopoietic growth
▪ used as signaling molecules in many cells of the body
INTERLEUKINS
HEMATOPOIETIC GROWTH
FACTOR
▪ regulating the proliferation and differentiation of HPCs as well
as regulating the survival and function of mature blood cells
▪ encoded by a single gene.
▪ erythropoietin - chromosome 7
▪ GM-CSF, IL-3, and M-CSF - long arm of chromosome 5.
▪ G-CSF - Chromosome 17
GENERAL CELLULAR
CHARACTERISTICS
1. Overall cell size
 SIZE  as AGE / MATURITY 
2. Nuclear-cytoplasmic ratio
 NUCLEAR SIZE  as AGE / MATURITY 
 NC ratio  as AGE / MATURITY 
GENERAL NUCLEAR
CHARACTERISTICS
1. Chromatin pattern
▪ LOOSE  CLUMPED
2. Nuclear shape
▪ very distinctive for particular cell types
3. Presence of nucleoli
▪ As cells mature, nucleoli are usually not visible.
CYTOPLASMIC
CHARACTERISTICS
1. Staining color and intensity
▪ Darker blue  lighter blue
2. Granulation
▪ None  non-specific  specific
3. Shape
▪ most distinctive variation in cytoplasmic shape occurs
in some blast forms, monocytes, and megakaryocytes.
CYTOPLASMIC CHARACTERISTICS

4. Quantity of cytoplasm
▪ Cytoplasm  as age 
5. Vacuolization
▪ Vacuoles  as age 
▪ Artifacts produce if the blood is stored for a longer-than-
acceptable period
6. Inclusion bodies
 TERMINOLOGIES
▪ RBCs are formally called erythrocytes.
▪ Nucleated precursors in the bone marrow are called erythroblasts
▪ Developing nucleated cells with normal appearance can be called
normoblasts
▪ Rubriblast terminology parallels the nomenclature used for
granulocyte development.
▪ Nuclei are always “baseball” round.
▪ As hemoglobin develops, the cytoplasm becomes more magenta.
PRONORMOBLAST / RUBRIBLAST/ PROERYTHROBLAST
▪ largest of the erythroid precursors
▪ the “mother cell”

 N:C ratio : 4:1

 Chromatin : fine and dark staining
 Nucleoli: 1 – 2
▪
oHeterogeneous
oDark blue (distinctively basophilic)
oNo granules
 Prorubricyte / Basophilic Normoblast (Erythroblast)

 N:C ratio : 4:1

 Chromatin : partially clumped, ‘wheel with broad spokes’.
 Nucleoli: ABSENT
▪
oDeeply basophilic
oNo granules
 Rubricyte / Polychromatic Normoblast (Erythroblast)
▪ “dawn of hemoglobinization”
▪ Last stage capable of mitosis

 N:C ratio : 1:1

 Chromatin : increasingly clumped
 Nucleoli: ABSENT
▪
omixtures of the red-staining of hemoglobin with the blue of RNA in
varying shades of gray
opink coloration mixed with basophilia  muddy, light gray
appearance
 Metarubricyte / Orthochromic Normoblast (Erythroblast)
▪ Nucleated RBC
▪ Mitosis no longer possible

 N:C ratio : LOW

Chromatin : pyknotic(dense or compact)
Nucleoli: ABSENT

oacidophilic (reddish pink) cytoplasm

osalmon-pink color
 Polychromatic Erythrocyte
▪ Called reticulocytes when stained with a vital stain
▪ No nucleus
▪ polychromatophilic as a result of the retention of RNA
▪ synthesizing hemoglobin, for 2 to 3 days in the marrow and for 1 day in
the blood
▪ seen in Wright’s stain as large bluish-red cells, polychromatophilic
macrocytes
 Erythrocyte
▪ No nucleus
▪ Biconcave disc
▪ salmon pink-staining cell with a central pale area
▪ area of central pallor of 1 to 3 μm ( 1/3 of the cell)
▪ circulate for about 120 days.
 ERYTHROCYTE

POLYCHROMATIC
ERYTHROCYTE

ORTHOCHROMATIC
NORMOBLAST

POLYCHROMATIC
NORMOBLAST

BASOPHILIC
NORMOBLAST

PRONORMOBLAST
GRANULOCYTIC SERIES
 MYELOBLAST
▪ earliest morphologically identifiable granulocytic precursor

 Shape : large oval to quadrangular nucleus

N:C ratio : 4:1
 Chromatin : very fine, uniform, finely reticular
Nucleoli: 1 – 5

osmall rim of basophilic cytoplasm that lacks granules, appears

smooth and velvety
 PROMYELOCYTE
▪ Production of azurophilic granules (NON-SPECIFIC GRANULES)

 Shape : oval and is often eccentric. A paranuclear halo or “hof”

 N:C ratio : 3:1
 Chromatin : more condensed than in the blast
 Nucleoli: 1

obasophilic and is filled by more and more azurophilic granules

opale grayish blue.
 MYELOCYTE
▪ “dawn of neutrophilia”
▪ Last stage capable of cell division

 Shape : Oval indented

 N:C ratio : 2:1
 Chromatin : red purple with slight granular, coarser, clumped

oappearance of secondary or specific cytoplasmic granulation

oNeutrophilic - fine and stain a blue-pink color
oEosinophilic – larger, oval shaped, orange with glassy texture
oBasophilic - dark blue-black color and a dense appearance
 METAMYELOCYTE
▪ Synthesis of tertiary granules
▪ color of the specific granulation continues to become a major
distinguishing feature

 Shape : indented or kidney bean shape (peanut shape),

 N:C ratio : 1:1
 Chromatin : more condensed or clumped

oPale blue to pinkish tan with moderate specific granules

 BAND / STAB

 Shape : Band shaped like a cigar band, C or S shaped, unable to see

filament,
 N:C ratio : 1:1
 Chromatin : coarsely clumped almost like leopard spot coarseness

oBrown-pink, with many fine secondary granules

 MATURE / SEGMENTED NEUTROPHIL

 Shape : Two to five lobes of nucleus connected by thin thread-like

filaments

oPale lilac with blue shading and many fine secondary dust-like
granules
oLife span: 6-10 hours
MATURE / SEGMENTED NEUTROPHIL
 EOSINOPHIL
▪ can appear at the myelocytic stages and move through the maturation
sequence.

 Shape : Eccentric nucleus, usually bilobed

oLarge, distinctive red-orange SPECIFIC granules with orange-pink

cytoplasm
oGranules are uniformly round, large, and individualized; if stain is
less than adequate, observe granules carefully for their crystalloid
nature
EOSINOPHIL
 BASOPHIL
▪ can appear at the myelocytic stages and move through the maturation
sequence.

 Shape : Coarse, clumped bilobed

oMany large SPECIFIC purple-black granules seem to obscure the large

cloverleaf form nucleus
omay decolorize during staining leaving pale areas within cell; granules
much larger than neutrophilic granules
BASOPHIL
MONONUCLEAR CELLS
 PROMONOCYTE
▪ earliest recognizable cell in this series

 Shape : oval or indented or folded

 N:C ratio : 3:1
 Chromatin : fine uniform or slightly streaked
 Nucleoli: 2-5

obasophilic with a ground-glass appearance and a variable number of

fine azurophilic granules
 MONOCYTE

 Shape : brainy convolutions to lobulated and S shaped,

horseshoe shaped,
 N:C ratio : 1:1
 Chromatin : loose-weaved, lacey, open, and thin

oAbundant gray-blue
oMay show area of protrusion or blebbing
oFine azure granules often referred to as azure dust or a
ground-glass appearance.
oCytoplasmic and nuclear vacuoles may also be present
 LYMPHOBLAST

 Shape : round
 N:C ratio : 4:1
 Chromatin : smudgy chromatin
 Nucleoli: 1-2 (surrounded by dark rim of chromatin)

oLittle, deep blue staining at edge

 PROLYMPHOCYTE

 Shape : round
 N:C ratio : 3:1
 Chromatin : slightly coarsened chromatin

oGray-blue, mostly blue at edges

 SMALL LYMPHOCYTE
▪ Clumping of chromatin around the nuclear membrane may help to
distinguish this from a nucleated cell
▪ Resting cell

 Shape : oval
 N:C ratio : 4:1
 Chromatin : coarse lumpy chromatin with specific areas of clumping,
a compact cell

oUsually just a thin border, with few azurophilic, red granules

 LARGE LYMPHOCYTE

 Shape : oval
 N:C ratio : 3:1
 Chromatin : Looser chromatin pattern, more
transparent

oCytoplasm is more abundant with tendency for

azurophilic granules
 LYMPHOCYTE DEVELOPMENT

▪ interleukin-1 (IL-1) and IL-6

▪ differentiate into the lymphoid stem cell
▪ bone marrow & thymus  primary or central lymphoid tissues
▪ lymph nodes spleen, and Peyer patches in the intestine  secondary
lymphoid tissue
▪ bursa of Fabricius, is present in birds and serves as the primary site of
B cell development
 B cell DIFFERENTIATION

▪ pro-B (progenitor) cell - first recognizable B cell in humans and

mammals
▪ CD19
▪ terminal deoxynucleotidyl transferase (TdT)
▪ contains no cytoplasmic or surface bound Ig
▪ pre-B cell
▪ characterized by the presence of intracytoplasmic μ heavy chains
with no surface-bound immunoglobulin
▪ virgin or naïve B cells
 B cell DIFFERENTIATION

▪ Germinal center B cells

▪ Bcl-6
▪ CD10
▪ Memory B cells
▪ CD27
▪ Either IgM or another heavy chain (rarely IgD alone)
▪ Plasma cells
▪ characterized by abundant cytoplasmic Ig
B cell DIFFERENTIATION
T cell DIFFERENTIATION & MATURATION

CD8
CLUSTER DESIGNATION
▪ CD 45  HSC
▪ CD 44  WBC
▪ Monocytes / macrophages
CD 16
CD 25
▪ NK cells
▪ CD 16
▪ CD 56
▪ CD 94
Mature B Cell
(After Blast Transformation)

 Shape : round or oval and may be eccentrically placed

Chromatin : fine pattern

onongranular,
omoderate in amount
omottled blue color
PLASMACYTOID LYMPHOCYTES

 Shape : round or oval and may be eccentrically placed

 Chromatin : coarse and irregularly spaced
 Nucleoli: may be visible

odistinctive dark blue with a lighter staining area, the hof,

next to the nucleus
PLASMA CELL

 Shape : small and eccentrically placed

 Chromatin : condensed and has a cartwheel configuration.
More than one nucleus may be seen

odark blue
othe hof area is usually visible
ooval in outline and abundant
oGranules are absent, but vacuoles are common
 OTHER FORMS OF PLASMA CELLS
▪ Grape or Mott cells
▪ cytoplasm is completely filled with Russell
bodies
▪ Flame cells
▪ the cytoplasm stains a bright-red color and
contains increased quantities of glycogen or
intracellular deposits of amorphous matter
MEGAKARYOCYTIC DEVELOPMENT

▪ Endomitosis
▪ nuclear division without cytoplasmic division
▪ burst-forming-unit megakaryocyte (BFU-M)
▪ most primitive progenitor cell committed to megakaryocyte
lineage
 MEGAKARYOBLAST
▪ most immature cell
▪ overlapping nuclear lobes and a small amount
of basophilic cytoplasm
▪ high nuclear to cytoplasmic ratio
▪ two to six nucleoli
▪ plasma membrane blebs
▪ blunt projections from the margin that resemble
platelets
 PROMEGAKARYOCYTE
▪ a large cell of 80 μm with dense alpha and lysosomal granules
▪ Nuclear lobularity first becomes apparent as an indentation
 MEGAKARYOCYTE
▪ largest bone marrow cells
▪ NC ratio : 1:12
▪ multilobular, not multinucleated
▪ nucleus is more compact,
▪ basophilia has disappeared
▪ granules are clustered into small aggregates
 PLATELETS
▪ no nucleus
▪ light blue, with evenly dispersed, fine red-purple granules
▪ life span of 7 to 10 days
▪ Formed by proplatelet process
▪ pseudopodial extensions of megakaryocytes that progressively
branch and thin out
▪ develop from invaginated surface membranes (demarcation
membranes)
REFERENCES
▪ Rodak’s Hematology Clinical Principles and Applications
▪ Ch 7 Hematopoiesis
▪ Ch 8 Erythrocyte Production and Destruction
▪ Ch 12 Leukocyte Development, Kinetics, and Functions
▪ Ch 13 Platelet Production, Structure, and Function

▪ Henry’s Clinical Diagnosis and Mgt. by Laboratory Methods

▪ Ch 31 Hematopoiesis

▪ Clinical Hematology: Theory and Procedures (Mary L. Turgeon)

▪ Ch 4 Hematopoiesis
▪ Ch 5 Erythrocyte Maturation, Physiology and Lifecycle
▪ Ch 14 Leukocytes : The Granulocytic and Monocytic Series
▪ Ch 16: Leukocytes: Lymphocytes and Plasma Cells
▪ Ch 23 Principles of Hemostasis and Thrombosis

▪ Hematology in Practice (Betty Ciesla)