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Blood, 15 December 2000, Vol. 96, No. 13, pp. 4169-4177
HEMATOPOIESIS
The  IIb 3 integrin and GPIb-V-IX
complex identify distinct stages in the maturation of CD34+
cord blood cells to megakaryocytes
Adeline Lepage,
Marylène Leboeuf,
Jean-Pierre Cazenave,
Corinne de la
Salle,
François Lanza, and
Georges Uzan
From INSERM U.506, Hôpital Paul Brousse, 14 avenue Paul Vaillant Couturier, F-94800 Villejuif, France; and INSERM
U.311, Etablissement Français du Sang-Alsace, 10 rue Spielmann,
B.P. 36, F-67065 Strasbourg Cedex, France.
Megakaryocytopoiesis is a complex multistep process involving cell
division, endoreplication, and maturation and resulting in the release
of platelets into the blood circulation. Megakaryocytes (MK)
progressively express lineage-restricted proteins, some of which play
essential roles in platelet physiology. Glycoprotein (GP)Ib-V-IX (CD42)
and GPIIb (CD41) are examples of MK-specific proteins having receptor
properties essential for platelet adhesion and aggregation. This study
defined the progressive expression of the GPIb-V-IX complex during in
vitro MK maturation and compared it to that of GPIIb, an early MK
marker. Human cord blood CD34+ progenitor cells were
cultured in the presence of cytokines inducing megakaryocytic
differentiation. GPIb-V-IX expression appeared at day 3 of culture and
was strictly dependent on MK cytokine induction, whereas GPIIb was
already present in immature CD34+ cells. Analysis by flow
cytometry and of the messenger RNA level both showed that GPV appeared
1 day later than GPIb-IX. Microscopy studies confirmed the late
appearance of GPV, which was principally localized in the cytoplasm
when GPIb-IX was found on the cell surface, suggesting a delayed
program of GPV synthesis and trafficking. Cell sorting studies
revealed that the CD41+GPV+ population
contained 4N and 8N cells at day 7, and was less effective than
CD41+GPV cells in generating burst-forming
units of erythrocytes or MK colonies. This study shows that the
subunits of the GPIb-V-IX complex represent unique surface markers of
MK maturation. The genes coding for GPIb-IX and GPV are useful tools to
study megakaryocytopoiesis and for tissue-specific or conditional
expression in mature MK and platelets.
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