Analysis of phorbol ester stimulated human megakaryocyte development
BJ Roth, GW Sledge , JE Straneva, J Brandt, M Goheen and R Hoffman
Department of Medicine, Indiana Elks Cancer Research Center, Indianapolis
46223.
Megakaryocytes are relatively rare components of human bone marrow, making
the study of their maturation difficult. Phorbol esters can act as
differentiating agents in a number of cell systems including murine
megakaryocytes. We report the effects of phorbol esters on the previously
described long-term human megakaryocytic leukemia cell culture, EST-IU.
While two nontransforming phorbols fail to affect these cells, the
transforming phorbol 12-O-tetradecanoylphorbol-13- acetate (TPA) induces a
phenotype with characteristics of more mature megakaryocytes in a
dose-related manner. This phenotype includes an increased adherence to
untreated plastic or glass, polyploidization, an increase in cell size, and
increased expression of both platelet glycoproteins and factor VIII-related
antigen. Two-color flow cytometric analysis allowed simultaneous
determinations of DNA content and the expression of surface membrane
antigens or alpha-granule constituents, providing evidence that nuclear,
membrane, and cytoplasmic maturation occur in parallel in this cellular
system. TPA- induced maturation of EST-IU cells provides an important new
cellular model for the further study of human megakaryocyte development.
Volume 72,
Issue 1,
pp. 202-207,
07/01/1988
Copyright © 1988 by The American Society of Hematology
