HEMATOPOIESIS

Ninik Sukartini
Department of Clinical Pathology
Faculty of Medicine, University of Indonesia

Modul Hematologi & Onkologi.

HEMATOPOIESIS

Prenatal, postnatal hematopoiesis

Medullary hematopoiesis,
extramedullary hematopoiesis

Effective hematopoiesis, ineffective

hematopoiesis
MEDULLARY HEMATOPOIESIS
 PRENATAL POSTNATAL

BIRTH

100
Yolk
per cent Bone
Sac
Liver marrow Vertebra

Sternum
Tibia
Spleen Rib
Femur
1 2 3 4 5 6 7 8 9 1 5 10 15 20 30 40 50 60 70

MONTHS AGE IN
YEARS
SITES OF HAEMOPOIETIC TISSUE IN FETUS AND
THROUGHOUT LIFE.
Fetus 0 2 months (yolk sac)
2 10 months (liver, spleen)
3 10 months (bone marrow)

Infants Bone marrow (practically all bones)

Adults Vertebrae, ribs, sternum, skull, sacrum and
pelvis proximal ends of femur
Hoffbrand AV. 2001
 Diagrammatic representation of bone marrow pluripotent
stem cells and the cell lines.
Hoffbrand AV. 2001
 T LYMPHOCYTE

LYMPHOID
B LYMPHOCYTE
STEM CELL

PLASMOCYTE
HEMATOPOIETIC
STEM CELL
ERYTHROCYTE

GRANULOCYTE
NONLYMPHOID
STEM CELL
MONOCYTE

THROMBOCYTE

Lineage of lymphoid and non lymphoid cells

Diggs Sturm Bell. 2005

 HEMATOPOIETIC GROWTH FACTORS
Hematopoietic growth factors are glycoprotein
hormones that regulated the proliferation &
differentiation of hemopoietic progenitor cells & the
function of mature blood cells

Control every step in the process of the development

of new blood cells

Stimulatory Inhibitory

Proliferation &
differentiation

Migration into
the circulation
Schiffman FJ. 1998 & Hoffbrand AV. 2001
 HAEMOPOIETIC GROWTH FACTORS
Act on stromal cells
IL-1
TNF

Act on pluripotential stem cells

Stem cell factor (SCF)
Flt ligand (Flt-L)

Act on multipotential progenitor cells G- and GM-CSF : granulocyte

IL-3 and granulocyte-macrophage
GM-CSF colony-stimulating factor;
IL-6
G-CSF IL : interleukin;
thrombopoietin M-CSF : macrophage colony-
stimulating factor;
Act on committed progenitor cells
G-CSF* TNF : tumour necrosis factor.
M-CSF
IL-5 (eosinophil-CSF) * These also act
erythropoietin synergistically with early
thrombopoietin* acting factors on pluripotential
progenitor.
Hoffbrand AV. 2001
THE ROLE OF GROWTH FACTORS IN NORMAL HAEMOPOIESIS

Hierarchical model of haemotopoiesis and lymphopoiesis including

important cytokines
Schiffman FJ. 1998.
 NOMENKLATUR
HEMATOPOIETIC TISSUES
CIRCULATING BLOOD
E. Myelocyte Eosinophil
Myeloblast Promyelocyte N. Myelocyte N. Metamyelocyte Neutrophi Neutrophil
B
l and Segmented
B. Myelocyte Basophil
Monoblast Promonocyte Monocyte
Stem cell

Megakaryoblast Promegakaryocyte Megakaryocyte Metamegakaryocyte Thrombocyte

Rubriblast Prorubricyte Rubricyte Metarubricyte Diffusely Erythrocyte
Basophilic
Erythrocyte

Lymphoblast Prolymphocyte Lymphocyte

Plasmoblast Proplasmocyte Plasmocyte

Blood cells

Diggs Sturm Bell. 2005

 MATURATION SEQUENCE OF RBC SYSTEM

MATURATION SEQUENCE
A. Cell size and cytoplasm
color
B. Nuclear size and color
C. Nuclear chromatin
structure
D. Composite (top to bottom :
rubriblast, prorubricyte,
rubricyte, metarubricyte,
diffusely basophilic
erythrocyte, erythrocyte

Diggs Sturm Bell. 2005

 MYELOCYTIC (GRANULOCYTIC) SYSTEM

A.Myeloblast
B.Promyelocyte
(progranulocyte)
C.Basophilic G.Neutrophilic K.Eosinophilic
myelocyte myelocyte myelocyte
D.Basophilic H.Neutrophilic L.Eosinophilic
metamyelocyte metamyelocyte metamyelocyte
E.Basophilic band I.Neutrophilic band M.Eosinophilic
F.Basophilic J.Neutrophilic band
segmented segment N.Eosinophilic
segmented

Diggs Sturm Bell. 2005

 NEUTROPHYL NON SEGMENTED AND SEGMENTED

MYELOCYTE METAMYELOCYTE BAND SEGMENTED

Terminology based on indentation of nuclei.

Nonsegmented

Segmented

Diggs Sturm Bell. 2005

 Cell types found on blood smears
from normal individuals
A Erythrocytes
B Large lymphocyte with
purplish-red (azurophil)
granules and deeply indented
by adjacent erythrocytes
C Neutrophilic segmented
D Eosinophilic segmented
E Neutrophilic segmented
F Monocyte with gray blue
cytoplasm, coarse linear
chromatin, blunt pseudopods
G Platelets (thrombocytes)
H Lymphocyte
I Neutrophilic band
J Basophil
Diggs Sturm Bell. 2005
 MONOCYTES

A.Monocyte with E.Monocyte with gray-

ground-glass blue color, band type
appearance evenly of nucleus, linear
distributed fine chromatin, blunt
granules, occasional pseudopods, and
azurophilic granules, granules
and vacuoles in F.Monocyte with gray-
cytoplasm blue color, irregular
B.Monocyte with opaque shape, and
cytoplasm and granules multilobulated nucleus
and with lobulation of G.Monocyte with
nucleus and linear segmented nucleus
chromatin H.Monocyte with
C.Monocyte with multiple blunt
prominent granules and nongranular
deeply indented pseudopods, nuclear
nucleus indentations, and folds
D.Monocyte without I.Monocyte with
nuclear indentations vacuoles and with
nongranular ectoplasm
and granular
endoplasm
Diggs Sturm Bell. 2005
 Maturation and differentiation of blood cells
Erythroid
enucleation Reticulocyte (supravital stain) erythrocyte

orthochromatic erythroblast

polychromatic erythroblast
proerythroblast basophilic erythroblast
Myeloid
segmented basophil
metamyelocyte band form

myelocyte
neutrophilic

eosinophilic
metamyelocyte band form segmented basophil
myeloblast I myeloblast II promyelocyte

basophilic

metamyelocyte
band form segmented basophil
Monocytic
monoblast tissue basophil
promonocyte

monocyte

macrophage

Yawata Y. 2001
 LYMPHOCYTES
A. Small mature H.Large lymphocyte
lymphocyte I.Large lymphocyte with
B. Lymphocyte of purplish-red
intermediate size (azurophilic) granules
C. Lymphocyte with J.Large lymphocyte with
indented nucleus irregular cytoplasmic
D. Lymphocyte of contour
intermediate size K.Large lymphocyte with
E. Lymphocyte with purplish-red
pointed cytoplasmic (azurophilic) granules
projections (frayed and with indentations
cytoplasm), typical caused by pressure of
nucleus erythrocytes
F. Spindle-shaped L.Large lymphocyte with
lymphocyte with purplish-red
indented nucleus (azurophilic) granules
G. Large lymphocyte
with indented
nucleus and
pointed cytoplasmic
projections
Diggs Sturm Bell. 2005
 DIFFERENTIATION AND CELL MARKERS OF LYMPHOCYTES
Pluripotent stem cell

CD 34+
Myeloid restricted
precursor

Lymphoid restricted
Tdt
precursor

CD34+
HLA DR
Tdt CD 34
CD 19
Pro-T cell Pro-B cell
Thymus
HLA DR
CD 19
CD 10
CD 34 Pre-B cell
Tdt
CD 7

Prothymocyte
m HLA DR
CD 19
CD 20
CD7 CD 22
CD2 C
CD5 Tdt D
CD4 3
CD8
Early thymocyte Pre-B cell
Antigen

IgM
CD7 CD7 HLA DR
CD3 TCR CD3 TCR CD 19
CD2 CD4 CD2 CD8
CD5 CD5 CD 20
CD 22
Mature T cell Mature T cell Mature B cell

Diggs Sturm Bell. 2005

Maturation and differentiation of non-lymphoid lineage (megakaryocyte)

megakaryocyte megakaryocyte

megakaryocyte

megakaryocyte

platelet

Maturation and differentiation of lymphoid lineage megakaryocyte

Natural killer lymphocyte

NK cell
Lymphoid cell of T-lineage
NK cell

mature T cell small lymphocyte (mature T cell)

T-intermediate
T-lymphoblast Lymphoid cell of B-lineage
lymphoblast

mature B cell small lymphocyte (mature B cell)

B-lymphoblast B-intermediate
lymphoblast

plasmocytoid
plasma cell
lymphocyte

Yawata Y. 2001
 MEGAKARYOCYTIC SYSTEM

A.Megakaryoblast with D.Metamegakaryocyte

single oval nucleus, with multiple nuclei
nucleoli, and bluish and with
foamy marginal thrombocytes
cytoplasmic (platelets)
structures E.Metamegakaryocyte
B.Promegakaryocyte nucleus with attached
with two nuclei, thrombocytes
granular blue F.Thrombocytes
cytoplasm, and (platelets)
marginal bubbly
cytoplasmic
structures
C.Megakaryocyte with
granular cytoplasm
and without discrete
thrombocytes
(platelets)
Diggs Sturm Bell. 2005
 A
Plasmocytic systems :
A. Plasmoblast
B. Proplasmocyte
B
C. Plasmocyte

C
Diggs Sturm Bell. 2005
 FIXED TISSUE CELLS
A.Tissue basophil (mast cell) B.Tissue eosinophil C.Tissue neutrophil
FIXED TISSUE CELLS
A.Phagocytic C. Tissue
histiocyte with neutrophil
vacuoles and with coarse
phagocytized nuclear
malarial pigment chromatin
B.Stem cell with structure,
partial rupture of neutrophilic
nuclear membrane granules, and
and blue nongranular shaggy margins
Diggs Sturm Bell. 2005 cytoplasm
 FAT CELLS
MACROPHAGES

A.Phagocytic histiocyte C.Phagocytic histiocyte,

with reticular fixed tissue type with
Top : fat cell with Bottom : fat cell
cytoplasmic structure, phagocytized hemosiderin
small round nucleus, showing cytoplasmic
vacuoles, and in cytoplasm
linear chromatin and lipoid bodies
phagocytized particles D.Ameboid phagocyte
globular body in separated by
B.Phagocytic histiocyte (wandering tissue
nucleus, ample reticular structures.
(ameboid macrophage) macrophage) with
cytoplasm with lipoid Structures
with phagocytized phagocytized particles
globules, winkled surrounding fat cells
erythrocytes and dark- and vacuoles in
membrane, reticular are mature
staining particles cytoplasm
stroma, and fibrillar erythrocytes.
Diggs Sturm Bell. 2005 marginal structures
 OSTEOBLASTS AND OSTEOCLAST

A.Osteoblast with C.Osteoclast large

prominent light multinucleated cell
zone in cytoplasm with uneven number
located away from of separated oval
nucleus nuclei, blue
B.Osteoblast with granules, and
oval eccentric frayed cytoplasmic
nucleus, distinct margins
linear chromatin
and nucleus, blue
bubbly cytoplasm
with prominent
light zone, and
fibrillar marginal
stuctures

Diggs Sturm Bell. 2005

 Blood oxygen
tension

Red cell Tissue (kidney) Phagocyte production Microbial invasion

production oxygen tension lymphocyte
differentiation
Inflammatory
Erythropoietin
mediators
Regulation of erythropoiesis. Regulation of white blood cell
production.

Vascular
integrity
Platelet production
Injury
Thrombopoietin

Bleeding
and hypoxia
Regulation of platelet
production.

Babior BM, Stossel TP. Hematology a pathophysiological approach. 1984.p.8-9.

 Bone marrow cells : normal adult values
Bone marrow cells Normal adult values (%)
Myeloblast 0 1
Promyelocyte 1 5
N. Myelocyte 2 10
N. metamyelocyte 5 15
Myeloid
N. Band 10 40
N. Segmented 10 30
Eosinophil 0 3
Basophil 0 1
Lymphocyte 5 15
Plasmocyte 0 1
Monocyte 0 2

Proerythroblast (rubriblast) 0 1
Basophilic erythroblast (prorubricyte) 1 4
Erythroid
Polychromatophilic erythroblast (rubricyte) 10 20
Orthochromatic erythroblast (metarubricyte) 5 - 10
Myeloid : erythroid (M : E) ratio (2 4) : 1

Diggs Sturm Bell. 2005

 Blood cells : normal adult values

Blood cells Percent Per uL

N. Band 2 6 100 600
N. Segmented 50 70 2.500 7.000
Eosinophil 1 3 50 300
Basophil 0 1 < 100
Lymphocyte 20 40 1.000 4.000
Monocyte 1 6 50 600

Total leukocyte counts : 5.000 10.000 / uL

Diggs Sturm Bell. 2005

EXTRAMEDULLARY HEMATOPOIESIS
EXTRAMEDULLARY HEMATOPOIESIS (EMH)
Is found in condition :
1. Excessive hematopoiesis
2. Bone marrow replacement
3. Dystrophic hematopoiesis
One or more hematopoietic cell lines may be present
EMH occur in :
1. Myeloproliferative disorders (MPD)
2. Myelodysplastic syndrome (MDS)
3. Acute leukemia
1. EMH EXCESSIVE HEMATOPOIESIS

Excessive hematopoiesis with blood forming cells

fill and expand the BM cavity, and eventually
extend beyond the bone to involve adjacent areas,
commonly paravertebral and epidural spaces
The liver, spleen and lymph nodes are also often
involved
Excessive hematopoiesis caused by :
1. Constitutional disorders : thalassemia, HbS
disease, hereditary spherocytosis
2. Acquired disorders : in neonatal periode,
immune hemolytic anemia, severe folate & vit
B12 deficiency
2. EMH BONE MARROW REPLACEMENT
Replacement of BM by non hematopoietic tissue
forces the distribution of hematopoiesis to other
sites commonly the liver, spleen and lymph nodes
BM occupied by histiocytes, granuloma, fibrous
tissue bone or malignant cell
PB shows leukoerythroblastosis : immature
granulocytes, nucleated red blood cell (NRBC)
Bone marrow replacement caused by :
1. Constitutional disorders : osteopetrosis,
Goucher disease
2. Acquired disorders : histoplasmosis,
tuberculosis, myeloma, metastatic carcinoma,
osteosarcoma
 3. EMH DYSTROPHIC HEMATOPOIESIS

Dystrophic hematopoiesis foci of disordered

growth, inflammation repair, and ossification

Dystrophic hematopoiesis caused by :

1. Tumours may show area of EMH as dystrophic

element

2. Result of tumours production of erythropoietin

EMH reported in the tumours of : hepatic tissue,

soft tissue, gonads uterus, central nervous system,
kidney and thyroid
Parhi DC, Chai CC, Edelman AS, Parveen T, Vo TLT. Pathology of bone marrow and blood cells.1st ed.
Philadelphia : Lippincott Williams & Wilkins; 2004.p.32-3.
INEFFECTIVE HEMATOPOIESIS
INEFFECTIVE HEMATOPOIESIS CHARACTERIZED BY :
Shortened blood cell survival
1. Usually macrocytic anemia
2. Reticulocyte count may be raised but is low
relative to the degree of anemia
3. indirect bilirubin serum
4. activity Lactate Dehydrogenase (LDH)
Morphologic abnormal blood cell precursors in the BM
A. Morphology abnormality of erythropoiesis
(dyserythropoiesis)
B. Morphology abnormality of granulopoiesis
(dysgranulopoiesis)
C. Morphology abnormality of thrombopoiesis
(dysthrombopoiesis)
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