title: Analysis of phorbol ester stimulated human megakaryocyte development.
abstract: 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.
mesh_terms: 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
chemical_list: 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