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Platelets generated in vitro from proplatelet-displaying human
megakaryocytes are functional
ES Choi, JL Nichol, MM Hokom, AC Hornkohl and P Hunt
AMGEN, Inc., Thousand Oaks, CA 91320.
An in vitro culture system demonstrating the transitions from megakaryocyte
progenitors to functional platelets is described. CD34- selected cells from
normal human peripheral blood are cultured under conditions that promote
megakaryocyte formation. After 8 to 11 days, enriched populations of mature
megakaryocytes are replated under conditions that favor the development of
proplatelets. Proplatelets express the platelet-specific proteins,
glycoproteins Ib and IIb (GPIb and GPIIb), and fibrinogen and also contain
microtubule coils equal in size to those found in plasma-derived platelets.
In addition, proplatelets have ultrastructural features in common with
plasma- derived platelets. Platelet-sized particles from the proplatelet
culture supernatants are examined. Ultrastructurally, these particles are
identical to plasma-derived platelets. Functionally, these culture- derived
platelets aggregate in response to both thrombin and adenosine diphosphate
(ADP) plus fibrinogen. This aggregation is specifically inhibited by the
addition of a function-blocking anti-GPIIbIIIa antibody. Culture-derived
platelets stimulated with agonists also express the activation-dependent
antigens P-selectin and functional fibrinogen receptor. This is the first
description of an in vitro culture system that sequentially demonstrates
megakaryocyte growth, development, and platelet production.
Volume 85,
Issue 2,
pp. 402-413,
01/15/1995
Copyright © 1995 by The American Society of Hematology
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