MEGAKARYOPOIESIS HEMATOLOGY II

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

BOOK TRANS: MEGAKARYOPOIESIS (STEININGER, RODAKS)

OVERVIEW MEGAKARYOCYTOPOIESIS
 Platelets
o Nonnucleated blood cells that circulate at 150 to 400
3 109/L.
o trigger primary hemostasis upon exposure to
subendothelial collagen or endothelial cell
inflammatory proteins at the time of blood vessel
injury.
 Wright-stained wedge-preparation blood film
o on this type of film, platelets are distributed
throughout the red blood cell monolayer at 7 to 21
cells per 1003 field.
o average diameter of 2.5 mm
 Megakaryocytes
o unique bone marrow cells where platelets arise.
o largest cells in the bone marrow
o Polyploid (possessing multiple chromosome copies)
 GATA-1 and FOG1 transcription slows, another
transcription factor, RUNX1, mediates the switch
from mitosis to endomitosis by suppressing the
Rho/ROCK signaling pathway, which suppresses
the assembly of the actin cytoskeleton.
 In response to the reduced Rho/ROCK signal,
inadequate levels of actin and myosin (muscle
fiber–like molecules) assemble in the cytoplasmic
constrictions where separation would otherwise
occur, preventing cytokinesis.
 Subsequently, under the influence of yet another
transcription factor, NF-E2, DNA replication
proceeds to the production of 8N, 16N, or even
32N ploidy with duplicated chromosome sets.
 Some megakaryocyte nuclei replicate five times,
reaching 128N; this level of ploidy is unusual (may
signal hematologic disease)

MUST KNOW NOTES: Wright-stained bone marrow

aspirate smear:
o each megakaryocyte is 30 to 50 mm in diameter with
a multilobulated nucleus and abundant granular
cytoplasm.
o microscopist may identify two to four megakaryocytes
per 103 low-power field.

MEGAKARYOCYTE DIFFERENTIATION AND PROGENITOR

MEGAKARYOCYTE PROGENITORS

 arise from the common myeloid progenitor under the

influence of the transcription gene product, GATA-1,
regulated by cofactor FOG1.
o Megakaryocyte differentiation is suppressed by
MYB
Fig. 1
 From the common myeloid progenitor there arise three
megakaryocyte lineage-committed progenitor stages:
o BFU-Meg - least mature burst-forming unit
o CFU-Meg - intermediate colony-forming unit
o LD-CFU-Meg - more mature progenitor light-density
CFU

Note: All three progenitor stages resemble lymphocytes and

cannot be distinguished by Wright-stained light microscopy
BFU-Meg and CFU-Meg
 Diploid and participate in normal mitosis, maintaining a
viable pool of megakaryocyte progenitors.

LD-CFU-Meg
 loses its capacity to divide but retains its DNA replication
MEGAKARYOCYTE DIFFERENTIATION AND PROGENITOR
and cytoplasmic maturation, a partially characterized form
 As endomitosis proceeds, megakaryocyte progenitors
of mitosis unique to megakaryocytes known as
leave the proliferative phase and enter terminal
endomitosis.
differentiation, a series of stages in which microscopists
become able to recognize their unique Wright-stained
LD-CFU-Meg
morphology in bone marrow aspirate films or hematoxylin
 is a form of mitosis that lacks telophase and cytokinesis
and eosin–stained bone marrow biopsy sections
(separation into daughter cells).

HEMA II MEGAKARYOPOIESIS Page 1 of 3

 MEGAKARYOPOIESIS
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3
 MK-I stage or megakaryoblast - least differentiated
megakaryocyte precursor
THROMBOCYTOPOIESIS (PLATELET SHEDDING)
Fig. 2 terminal megakaryocyte shedding
 One cannot find reliable evidence for platelet budding or
shedding simply by examining megakaryocytes in situ,
even well-structured bone marrow biopsy preparations.
However, in megakaryocyte cultures examined by
transmission electron microscopy (TEM):
o Demarcation system (DMS) dilates
o Longitudinal bundles of tubules form
o Proplatelet processes develop
o Transverse constrictions appear throughout the
proplatelet processes
 In the bone marrow environment, processes are believed
to pierce through or between sinusoid lining endothelial
cells, extend into the venous blood, and shed platelets.
Fig. 3
HEMA II MEGAKARYOPOIESIS
HEMATOLOGY II
MEGAKARYOCYTE MEMBRANE RECEPTOR AND MARKERS
 In specialty and tertiary care laboratories, scientists and
technicians employ the following to identify visually
indistinguishable megakaryocyte progenitors in
hematologic disease:
o Immunostaining of fixed tissue
o Flow cytometry with immunologic probes
o Fluorescent in situ hybridization (FISH) with genetic
probes
Fig. 4
HORMONES AND CYTOKINES OF MEGAKARYOCYTOPOIESIS
 Messenger ribonucleic acid (mRNA) for TPO has been
found in the:
o Kidney
o Liver- has the most copies and considered the
primary source
o Stromal cells
o Smooth muscle cells
TPO (THROMBOPOIETIN)
 circulates as hormone in plasma
 ligand that binds the megakaryocyte and platelet
membrane receptor protein, MPL, named for v-mpl, a viral
oncogene associated with murine myeloproliferative
leukemia.
 Induces the proliferation and maturation of
megakaryocytes
 Induces megakaryopoiesis, or platelet release
 Using in vivo and in vitro experiments, TPO, in synergy
with cytokines:
o Induces stem cells to differentiate into megakaryocyte
progenitor
o Further induces the differentiation of megakaryocyte
progenitors into megakaryoblasts and
megakaryocytes.
 The plasma concentration of TPO is inversely
proportional to platelet and megakaryocyte mass,
implying the membrane binding and consequent removal
of TPO by platelets is the primary platelet count
mechanism.
 Synthetic TPO mimetics (analogues) - elevate the
platelet count in patient being treated for variety of
cancers, including acute leukemia.
Page 2 of 3
 MEGAKARYOPOIESIS HEMATOLOGY II
Luyaben, Pilit, Baligod, Cabucana, Dangelan Topic# 3

RECEPTOR MPL AGONIST  Metamegakaryocyte


TM
Romiplostim (NPlate , Amgen Inc., Thousand Oaks, - very large cell, many times the size of a mature
CA, FDA cleared in 2008) granulocyte, with a decreased nuclear-cytoplasmic
o a commercial one ratio compared with the immature stages of
o a nonimmunogenic oligopeptide that is also development.
effective in raising the platelet count in immune
thrombocytopenic purpura BOOK REFERENCES/PPT:
Maam Donna therese Taguinod PPT: Topic 5

® ®
Eltrombopag (Promacta and Revolade , Glaxo Smith
Megakaryopoiesis
Kilne, Inc., Philadelphia, PA, FDA cleared in 2011) th
Rodaks Hematology 5 Edition
o a second nonpeptide MPL receptor agonist Steininger Clinical Hematology
o binds and activate an MPL site separate from
romiplostim.
o They may have additive effects.

OTHER CYTOKINES THAT FUNCTION WITH TPO TO STIMULATE

MEGAKARYOPOIESIS
 Interleukin 3 (IL-3) - Act in synergy with TPO to induce
the early differentiation of stem cells
 Interleukin 6 (IL-6)
 Interleukin 11 (IL-11)
®
o Oprelvekin (Neumega , Wyeth Ayerst Genetics
Institute, Cambrigde, MA, FDA cleared in 1997)
– An IL-11 polypeptide mimetic
– Stimulates platelet production in patients
withchemotheraphy-induced
thrombocytopenia

IL-6 and IL-11 act in the presence of TPO to enhance the

later phenomena of endomitosis, megakaryocyte maturation
and thrombocytopoiesis.

Additional Notes Only from Steininger Book

Megakaryocyte Maturation
 Megakaryoblast- Earliest recognizable state of
maturation
Features:
o Displays blunt protrusions from is cytoplasmic
membrane
o Central area: Contains mitochondria and a primitive
endoplasmic reticulum
o Cytoplasm: has alpha granules and centrioles
 James Wright – provided evidence that platelets where
derived from megakaryocytes within the bone marrow.
 Thrombopoietin – a specific hormone that is responsible
for the commitment of the megakaryoblast to differentiate
further into more mature stages.
 Promegacaryocyte
- Megakaryoblast, as it matures to the
promegakaryocyte stage increases in volume
 Megakaryocyte
- Known as the stage that does not ordinarily produce
platelets.
- However, megakaryocytes with at least 4 nuclei can
produce platelets.

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