MEGAKARYOPOIESIS HEMATOLOGY II

Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3

TRANS PPT TOPIC 4: PLATELET DEVELOPMENT hematopoiesis with a decline in

primitive hematopoiesis of the yolk
TOPIC OUTLINE sac.
I. Overview on Hematopoiesis
II.
 Begins prior to the 5th month of
gestation and remain as the chief
site at birth.
MEDULLARY  During this phase the HSCs and
(MYELOID) mesenchymal cells migrate into the
PHASE Phase core of the bone.
 Measurable levels of erythropoietin
(EPO), granulocyte colony-
stimulating factor (G-CSF),
granulocyte macrophage colony-
stimulating factor (GM-CSF), and
hemoglobins F and A can be
detected.

OVERVIEW ON HEMATOPOIESIS  PRODUCTION OF SPECIFIC CELLS

 HEMATOPOEISIS o Granulopoiesis: 2nd month of gestation
o It is a continuous, regulated process of blood cell o Megakaryopoiesis: 2nd month of gestation
production that includes cell renewal, proliferation, o Lymphopoiesis: 4th month
differentiation, and maturation. o Monopoiesis: 5th month
o A hematopoietic stem cell is capable of self-renewal
and directed differentiation into all required cell
lineages.
o During fetal development, the restricted, sequential
distribution of cells initiates in the yolk sac and then
progresses in the aorta-gonad mesonephros (agm)
region (mesoblastic phase), then to the fetal liver
(hepatic phase), and finally resides in the bone
marrow (medullary phase).
o In healthy adults hematopoiesis is restricted
primarily to the bone marrow.
 STAGES OF INTRAUTERINE HEMATOPOIESIS
o Mesoblastic Phase
o Hepatic Phase
o Medullary (Myeloid) Phase
STAGES OF INTRAUTERINE HEMATOPOIESIS Fig. 1 Site of Hematopoiesis by Age

 Primitive erythroblast
ADULT HEMATOPOIETIC TISSUES
- important in early
embryogenesis to produce
hemoglobin. PRIMARY LYMPHOID SECONDARY LYMPHOID
MESOBLASTI  Gower 1 ORGAN ORGAN
C PHASE  Gower 2 This is antigen-independent Antigen-dependent
 Portland lymphopoiesis. lymphopoiesis
 Aorta-gonad-mesonephros region  Thymus gland  Lymph nodes
o site of pre-T cell  Spleen
- give rise to hematopoietic stem
cells for definitive o permanent differentiation  GALT Gut Associated
adult hematopoiesis.  Bursa Fabricus (Bone LymphoidTissue
marrow (tonsils,Peyer'sPatches)
 Hematopoiesis begins at 5 to 7
o Site of pre-B cell
gestational weeks
differentiation;
 Liver is the major site of
HEPATIC o Responsible for
hematopoiesis during the second
PHASE trimester of fetal life. normal antibody
 The developing erythroblasts signal production
the beginning of definitive

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 MEGAKARYOPOIESIS HEMATOLOGY II
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3

ADULT HEMATOPOIETIC TISSUES

HEMATOPOIETIC STEM CELL AND CYTOKINES

RED MARROW YELLOW MARROW

 Hematopoietically active
 Marrow consisting of the
developing blood cells
and their progenitors.
 Hematopoietically
inactive Marrow
composed primarily of
adipocytes (fat cells),
with undifferentiated
mesenchymal cells and
macrophages.

DEFINITION OF TERMS

 Marrow Cellularity - the ratio of red marrow cells to fat

(red marrow/ yellow marrow)
o Normocellular: marrow has 30-70% hematopoietic
cells
o Hypercellular/hyperplastic: marrow has >70%
hematopoietic cells
o Hypocellular/hypoplastic: marrow has <30%
hematopoietic cells
o Aplastic: marrow has few or no hematopoietic cells
 M:E (Mveloid:Ervthroid) ratio - the ratio of granulocytes
and their precursors to nucleated erythroid precursors.
 OTHER HEMATOPOIETIC TISSUES:
o SPLEEN
o LIVER
o LYMPH NODES
o THYMUS
Fig 3. Hematopoietic Stem Cell

 TILL AND MCCULLOCH

o Proposed that hematopoiesis is a random process
where by hematopoietic stem cell randomly commit to
self renewal or differentiation.
o This model is also called stochastic model of
hematopoiesis.
 When hematopoietic Stem Cell Divides it gives Rise to 2
identical daughter cells
o Symmetric division
o Asymmetric division
o Apoptosis

Fig. 2 Other Hematopoietic Tissues

Fig. 4

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 MEGAKARYOPOIESIS
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3
MEGAKARYOPOIESIS
Fig. 5 Thrombopoietin
 THROMBOPOIETIN (TPO)
MEGAKARYOCYTES MEMBRANE RECEPTOR AND MARKERS
o It is a 353-amino acid protein located on chromosome
3p27.
o 70 KD molecule with 23% homology to EPO
o Produced in the liver, kidneys and smooth muscles
o Ligand that binds megakaryocytes and platelet
membrane receptor protein (C-MPL)
o Induces stem cell differentiation
Fig. 6 Endomitosis
 PLOIDY- number of chromosomes occurring in the nucleus of a
cell
HEMA II INTRO TO CLINICAL CHEM (LEC#1)
HEMATOLOGY II
Fig. 7 Terminal megakaryocyte Differentiation
 There are several flow cytometric megakaryocyte
membrane markers, including MPL, which is the TPO
receptor site present at all maturation stages, and the
stem cell Intracellular and common myeloid progenitor
marker CD34.
 The CD34 marker disappears as differentiation proceeds.
Fig. 9 Megakaryocyte Receptor
 The platelet membrane glycoprotein llb/lla (CD41, a
marker located on the llb portion) first appears on
megakaryocyte progenitors and remains present
throughout maturation, alon8 with immunologic markers
CD36, CD42, CD61, and CD62.
 Cytoplasmic coagulation factor VIl, von Willebrand tactor
(VWF), and Fibrinogen, may be detected immunostaining
in the fully developed megakaryocyte.
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 MEGAKARYOPOIESIS HEMATOLOGY II
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3

HORMONES AND CYTOKINES OF MEGAKARYOPOIESIS DIAMETER 15-50 um

 The growth factor TPO is a 70,000 Dalton molecule that CYTOPLASMIC Absent in deep blue cytoplasm
possesses 23% homology, with the red blood cell GRANULES
producing hormone erythropoietin. CYTOPLASMIC TAGS Present (blunt cytoplasmic
(Protusion) protrusion)
MEGAKARYOCYTE
NUCLEAR FEATURES Single nucleus, visible nucleoli;
 Other cytokines and hormones that participate Centrally located w/ fine delicate
synergistically with TPO and the interleukins (interleukin-3 chromatin strands
(|L-3), IL-6, and IL-11) are: VISIBLE PLATELETS No
o include stem cell factor, also called kit ligand or mast
cell growth factor; DIAMETER 20-80 um
o granulocyte-macrophage colony-stimulating fac- tor CYTOPLASMIC Few in Blue cytoplasm
GM-CSF); GRANULES
o granulocyte colony-stimulating factor (G-CSF); CYTOPLASMIC TAGS Present
SUBSTANCE THAT INHIBIT MEGAKARYOCYTE GROWTH (Protusion)
o acetylcholinesterase-derived megakaryocyte growth NUCLEAR FEATURES Double nucleus (2N), Fine
stimulating peptide and. chromatin, Visible Nucleoli
 Platelet factor 4 (PF4) VISIBLE PLATELETS No
 Formation of a network:
METAMEGAKARYOCYTE o DEMARCATING MEMBRANE SYSTEM (DMS) -
 B-thromboglobulin future membrane of the metamegakaryocyte's new
 Neutrophil- activating peptide 2 offspring - the platelets
 IL-8 and other factors inhibit in vitro megakaryocyte growth
which indicates that they may have a role in the control of CYTOPLASMIC Numerous, small redddish granules in
megakaryopoiesis in vivo GRANULES abundant, pinkish-purple cytoplasm
Reduction in the transcription factors FOG1, GATA-1, CYTOPLASMIC Usually absent
and NF-E2 diminish megakaryopoiesis at the TAGS
progenitor, endomitosis, and terminal maturation NUCLEAR 2 or more nuclei (4N to 8N)-
phases. FEATURES endomitosis; coarse chromatin
VISIBLE NO except in myeloproliferative d/o,
PLATELETS myelophthisic process
 APPROXIMATELY 5 DAYS MATURATION IN THE BM
o AT LEAST 4 NUCLEUS COULD PRODUCE
PLATELETS

MEGAKARYOBLAST

THROMBOPOEISIS
Fig. 10 Hormones and Cytokines that Control Megakaryopoiesis
CYTO Aggregated
PLAS
MIC
GRAN
ULES
CYTO Absent
PLAS
MIC
TAGS
NUCL
MEGAKARYOBLAST 4 or more nuclei
EAR (4N to 32N)
FEAT
URES
VISIB YES
LE
PLAT
ELET
Fig. 11 Thrombopoeisis S
 Growth Factors: MEG-CSF AND THROMBOPOIETIN  Stage of cytoplasmic division
 1 metamegakaryocyte = 1000 - 4000 platelets

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 MEGAKARYOPOIESIS HEMATOLOGY II
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3

 9-12 days lifespan of cytoplasmic fragments of

megakaryocytes

INITIATION OF PLATELET FORMATION

1. DMS (demarcation system) dilates
2. Longitudinal bundles of tubules are formed
3. Proplatelet process extends (pseudopods)
4. Transverse constriction
5. Proplatelet process pierces through the sinusoidal lining
of endothelial cells
6. Extends into the venous blood
7. Releases platelet

Fig 12. Platelet

KEY POINTS
Thrombopoiesis/Megakaryopoiesis:
- Platelet Maturation Series (starting from the youngest
to the most mature form)
1. Cell size increases with maturation
2. Repeated nuclear division occurs without cellular
division ( endomitosis) The nucleus is polyploid which
means that the nucleus contain an increased number
of chromosomes over the normal diploid number ( 2N
to 64N)
3. The final mature functional form is a cytoplasmic
fragment (platelet)

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