Analysis of phorbol ester stimulated human megakaryocyte development.
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.
D000941:Antigens / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D005165:Factor V / Q000378:metabolism; D005169:Factor VIII / Q000276:immunology / Q000378:metabolism; D006410:Hematopoiesis / Q000187:drug effects*; D006801:Humans; D008533:Megakaryocytes / Q000187:drug effects* / Q000648:ultrastructure; D010703:Phorbol Esters / Q000494:pharmacology*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011003:Ploidies; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014841:von Willebrand Factor
D000941:Antigens; D010703:Phorbol Esters; D010980:Platelet Membrane Glycoproteins; D014841:von Willebrand Factor; C008876:phorbol-12,13-didecanoate; C028114:croton factor F2; D005165:Factor V; D005169:Factor VIII; D013755:Tetradecanoylphorbol Acetate