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Culture of isolated bovine megakaryocytes on reconstituted basement
membrane matrix leads to proplatelet process formation
KS Topp, F Tablin and J Levin
Department of Anatomy, School of Veterinary Medicine, University of
California, Davis 95616.
We have enriched for bovine megakaryocytes and identified a culture system
that may provide an in vitro model for platelet formation. Mature
megakaryocytes with an unusually high ploidy distribution were obtained
after differential centrifugation and velocity sedimentation of bone marrow
cells through gradients of bovine serum albumin (BSA). The cell membranes
of isolated megakaryocytes and megakaryocytes in vivo stained with antisera
to human platelets and human platelet membrane GPIIIa. The microenvironment
of bovine megakaryocytes in vivo was investigated using antibodies to types
I and IV collagen and laminin. In an attempt to duplicate the
microenvironment in vitro, bovine megakaryocytes were cultured on a
reconstituted basement membrane matrix (Matrigel). The cells adhered to the
gel, extended radial lamellipodia, and occasionally formed lengthy
pseudopodia. Ultrastructural examination of these cells showed widening and
coalescence of the megakaryocyte demarcation membranes (DMS), and inclusion
of platelet granules, thin filaments, and microtubules in the processes.
Very few DMS vesicles were present distally in the processes. The culture
of megakaryocytes on a reconstituted basement membrane may closely model
the in vivo megakaryocyte microenvironment and allow the study of
thrombocytopoiesis in vitro.
Volume 76,
Issue 5,
pp. 912-924,
09/01/1990
Copyright © 1990 by The American Society of Hematology
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