THROMBOCYTE

By: Yoavita
Moderator: dr.C.L. Retno H., Sp.PK
INTRODUCTION
• Platelets were recognized in 1882 by Bizzozero as a cell
structure different from red and white cells
• In 1970, platelets’ relationship to hemostasis and
thrombosis became so important
• Every cubic millimeter of blood contains one-fourth of 1
billion platelets, resulting in approximately a trillion
platelets in the blood of an average woman
• Each platelet makes 14,000 trips through the
bloodstream in its life span of 7 to 10 day
MEGAKARYOCYTOPOIESIS
MEGAKARYOCYTOPOIESIS
• Megakaryocytes (MKs) give rise to circulating platelets
(thrombocytes) through commitment of the multipotent stem
cell to the MK lineage, proliferation of the progenitors and
terminal differentiation of MK

• This process is characterised by DNA endoreduplication,

cytoplasmic maturation and expansion, and release of
cytoplasmic fragments as circulating platelet
MEGAKARYOCYTOPOIESIS
MEGAKARYOCYTOPOIESIS
Megakaryocyte Progenitors

• The hematopoietic stem cell, CFU-GEMM, differentiates to

the megakaryocyte lineage under the influence of hormon
thrombopoietin (TPO) and a series of cytokines

• 3 megakaryocyte lineage-commited progenitor stages:

1. BFU-Meg
2. CFU-Meg
3. LD-CFU-Meg
MEGAKARYOCYTOPOIESIS
Terminal Megakaryocyte Differentiation

• Megakaryocyte progenitors leave the proliferative phase and

enter terminal differentiation

• Past the LD-CFU-Meg there is no further mitosis, but three to

four morphologically identifiable stages

MK I Megakaryoblast
MK II Promegakaryocyte
MK III Megakaryocyte
MK IV Postmature  multilobed, highly condensed nucleus
MEGAKARYOCYTOPOIESIS
Megakaryocyte Maturation

• Megakaryocyte maturation is marked by a mysterious form of

mitosis that lacks telophase and cytokinesis
 endoreduplication or endomitosis

• DNA synthesis proceeds to the production of 8N, 16N, or 32N

ploidy with completely duplicated sets of chromosomes but
no cell division
MEGAKARYOCYTOPOIESIS
Megakaryocyte Maturation
MEGAKARYOCYTOPOIESIS
Megakaryocyte Maturation
MK I MK II MK III
% Precursors 19 25 56
Diameter 14 – 18 μm 15 – 40 μm 30 – 50 μm
Nucleus Round Indented Multilobed
Nucleoli 2–6 Variable Variable
Chromatin Homogenous condensed Deeply condensed
N:C ratio 3:1 1:2 1:4
MEGAKARYOCYTOPOIESIS
Megakaryocyte Maturation
MK I MK II MK III
Mitosis Present Absent Absent
Cytoplasm Basophilic Basophilic and Eosinophilic and
granular granular
α-granules Present Present Present
δ-granules Present Present Present
DMS Present Present Present
MEGAKARYOCYTOPOIESIS
Thrombopoiesis

• As the MK matures, the polyploid nucleus becomes

horseshoe-shaped, the cytoplasm expands, and platelet
organelles and the demarcation membrane system are
amplified

• The robust cytoplasmic mass forms proplatelet projections

which give rise to de novo circulating platelet
MEGAKARYOCYTOPOIESIS
Regulation of Megakaryocytopoiesis
TPO IL-11 IL-3 IL-6
Differentiation to
- 0 + 0
progenitors
Differentiation to
+ + + 0
megakaryocytes
Late maturation + + 0 +
Thrombopoiesis 0 + - +
Clinical use Trials trials - -
MEGAKARYOCYTOPOIESIS
Expression of cell surface markers
PLATELETS
PLATELETS
• The propetelet process shed platelets, cells consisting of
granular cytoplasm with a membrane but no nuclear material,
into the venous sinus

• The diameter averages 2.5 μm

PLATELET ULTRASTRUCTURE
• Platelets, although anucleate, are strikingly complex and
metabolically active

• Their ultrastructure has been studied using scanning and

transmission electron microscopy, flow cytometry, and
molecular sequencing techniques
PLATELET ULTRASTRUCTURE
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane

• Resembles any biologic membrane: a bilayer composed of

proteins and lipids

• The predominant lipids are:

─ Phospholipid  form the basic structure (a bilayer)
─ Cholesterol  stabilizes the membrane, maintais fluidity,
and help control the transmembranous
passage of materials
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane

• Anchored within the membrane are glycoproteins and

proteoglycans which support surface glycosaminoglycans,
oligosaccharides, and glycolipids

• The platelet membrane surface, the glycocalyx, also absorbs

albumin, fibrinogen, and other plasma proteins, in many
instances transporting them to storage organelles by
endocytosis
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane

• The plasma membrane is selectively permeable

• The membrane bilayer provides phospholipids that support

platelet activation internally and plasma coagulation
externally

• The glycoproteins support essential plasma surface-oriented

glycosylated receptors that respond to cellular and humoral
stimuli, integrating their stimulus with internal organelles
PLATELET ULTRASTRUCTURE
Surface-Connected Canalicular System (SCCS)

• The plasma membrane invaeds the platelet interior, producing

its uniques SCCS

• The SCCS twists spongelike throughout the platelet, enabling

platelet to store additional quantities of the hemostatic
proteins of the glycocalyx

• The SCCS is the route for endocytosis and for secretion of

granular contents on activation
PLATELET ULTRASTRUCTURE
Dense Tubular System (DTS)

• A condensed remnant of the rough endoplasmic reticulum

• The DTS sequesters Ca2+ and bears a series of enzymes that

support platelet activation: phospholipase A2, cyclo-
oxygenase, and thromboxane synthase

• The DTS is the control center of platelet activation

PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane Receptor for Adhesion
Nomeclature Ligand CD
GP Ia/IIa Integrin: α2β1 Collagen CD49b, CD29
GP Ic/IIa Integrin: α5β1 Laminin CD49e, CD29
GP Ic/IIa Integrin: α6β1 Fibronectin CD49f, CD29
GP IV Miscellanous platelet Collagen II and
CD36
receptor thrombospondin
GP VI CAM of the Ig gene
Collagen -
family
GP Ib/IX/V CAM of the leucin-rich- vWF and CD42b, CD42c
repeat family thrombin CD42a, CD42d
GP IIb/IIa Integrin: αIIbβ3 Fibrinogen, vWF CD41, CD61
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane Receptor for Aggregation
STR-Ligand interaction coupled to
Ligand
signaling G-proteins
PAR1 Thrombin Coupled to G1 protein that reduces cAMP;
Coupled to Gq and G12 proteins that
increase IP3 and DAG
PAR4 Thrombin Coupled to Gq and G12 proteins that
increase IP3 and DAG
P2Y1 ADP Coupled to Gq protein that increase IP3
and DAG
P2Y12 ADP Coupled to G1 protein that reduces Camp
PLATELET ULTRASTRUCTURE
Platelet Plasma Membrane Receptor for Aggregation
STR-Ligand interaction coupled to
Ligand
signaling G-proteins
TPα & TPβ TXA2 Coupled to Gq protein that increase IP3
and DAG
α2-adrenergic Epinephrine Coupled to G1 protein that reduces cAMP;
potentiates effects of ADP, thrombin, and
TXA2
IP PGI2 Coupled to Gs protein that increases cAMP
to inhibit activation
PLATELET ULTRASTRUCTURE
Additional Platelet Membrane Receptors
ICAMs CD50, CD54, Play a role in inflammation and the immune
CD102 reaction
PECAM CD31 Mediates platelet to WBC and platelet to
endothelial cell adhesion
FcγRIIA CD32 A low affinity receptor for the Ig Fc portion
that plays a role in heparin-induced throm-
bocytopenia
P-selectin CD62 Encourages plaelets to bind endothelial cells,
leukocytes, and each other
PLATELET ULTRASTRUCTURE
Platelet Cytoskeleton

• A thick circumferential bundle of microtubules maintains the

platelet’s shape

• In the narrow area between microtubules and membrane lies

a thick meshwork of microfilaments composed of actin

• Actin is contractile in platelets and anchors the plasma

membrane glycoproteins and proteoglycans
PLATELET ULTRASTRUCTURE
Platelet Cytoskeleton

• The cytoplasm also contains intermediate filaments, ropelike

polymers 8 to 12 ηm in diameter, of desmin and vimentin

• Microtubules, actin, microfilaments, and intermediate

microfilaments control platelet shape change, extension of
pseudopods, and secretion of granule contents
PLATELET ULTRASTRUCTURE
Platelet Granules

Small molecules
ADP
ATP
Calcium
Serotonin

Large molecules
β-thromboglobulin
Factor V, XI
Protein S β-N-acetylglucosaminidase
Fibrinogen β -N-acetylgalactosaminidase
Vwf β -Glucoronidase
Platelet factor 4 β -Galactosidase
PDGF α-Arabinosidase
PLATELET ACTIVATION
Adhesion

• Platelets repair minor injuries to blood vessel linings, such as

regular sloughing of senescent endothelial cells, through
adhesion

• Platelets reversibly bind elements on the vascular matrix

• In veins or venules, platelets may adhere directly or through

CAMs GP Ia/IIa, GP IV, and GP VI
PLATELET ACTIVATION
Adhesion

• In capillaries and arterioles, platelets adhere to fibrillar vWF

through their GP Ib/IX/V receptor

• Adhesion require actin contraction and the formation of

pseudopods
PLATELET ACTIVATION
Aggregation

• When vessel damage is extensive, platelets adhere and

aggregate

• Platelets irreversibly bind each other

• Aggregation requires the conformation of the GP IIb/IIIa

integrin and includes pseudopod formation and redistribution
of P-selectin to the surface membrane
PLATELET ACTIVATION
Secretion

• “outside-in” platelet activation through ligand binding to

integrins and STRs triggers actin microfilament contraction

• Intermediate filaments and microtubules contract,

compressing granules

• Contents of α-granules and lysosomes flow through the SCCS

PLATELET ACTIVATION
Secretion

• Contents of δ-granules are secreted through the plasma

membrane

• The α-granule contents are vasoconstrictors and platelet

agonists that amplify primary hemostasis; several of the α-
granule contents are coagulation proteins
PLATELET ACTIVATION
PLATELET DISORDERS
PLATELET DISORDERS
Disorders

Quantitative Qualitative

Thrombocytopenia Hereditary

Thrombocytosis Acquired
PLATELET DISORDERS
Quantitative
Thrombocytopenia
Platelets < 100,000/μL
Impaired/decreased production
 Bernard Soulier syndrome
 Fanconi anemia
 Viral infection
 Drug-induced
Increased destruction
 Immune: ITP, PTP, etc
 Non-immune: TTP, HUS, DIC, etc
Related to distribution/dilution
 Splenic sequestration
 Massive blood transfusions
 Kasabach-Merritt syndrome
Thrombocytosis
Platelets > 450,000/μL
Primary
 Polycythemia vera
 Chronic myelogenous leukemia
 Myelofibrosis & myeloid metaplasia
 Essential thrombocythemia
Secondary
 Blood loss
 Chronic inflammatory diseases
 Postsplenectomy
 Iron deficiency anemia
 Stress & exercise
PLATELET DISORDERS
Qualititative
Hereditary
Disorders of platelet adhesion
 von Willebrand disease
 Bernard Soulier syndrome
Disorders of primary aggregation
 Glanzmann thrombasthenia
 Hereditary afibrinogenemia
Disorders of platelet secretion
 Storage pool disease
 Chédiak-Higashi syndrome
 Herman-Pudlak syndrome
 Wiskott-Aldrich syndrome
Acquired
Disorders of platelet adhesion
 Chronic liver disease
 Myeloproliferative and
lymphoprolifera-tive disorders
Disorders of primary aggregation
 Acquired von Willebrand disease
 Uremia
Disorders of platelet secretion
 Drug-induced
REFERENCES
1. Ciesla B, editor. Hematology in Practice. Philadelphia: FA
Davis. 2007.
2. Rodak BF, Fritsma GA, Doig K, editors. Hematology: Clinical
Principles and Applications, 3rd ed. Missouri: Saunders
Elsevier. 2007.
3. Lichtman MA, Beutler E, Seligsohn U, Kaushansky K, Kipps
TO, editors. Williams Hematology, 7th ed. New York:
McGraw-Hill. 2005.
THANK YOU