The role of erythropoietin, thrombopoietic stimulating factor, and myeloid colony-stimulating factors on murine megakaryocyte colony formation.

Various growth factors including purified erythropoietin, colony-stimulating factor (CSF-1), and granulocyte-macrophage CSF (GM-CSF) were tested for their ability to stimulate megakaryocytopoiesis. Four separate preparations of erythropoietin were tested in highly defined cell culture medium. One unit of purified material stimulated small but significant numbers of megakaryocyte colonies, both in serum-containing and in serum-free cultures. All other erythropoietin preparations failed to induce megakaryocyte colony formation. Purified erythropoietin showed no synergistic activity with either WEHI-3 cell conditioned medium (WEHI-3CM, a source of both megakaryocyte CSF and megakaryocyte-potentiating activity) or P388D1 cell conditioned medium (P388D1CM, a preparation containing megakaryocyte potentiator). Partially purified thrombopoietic stimulatory factor did not stimulate directly megakaryocyte colony formation, but acted together with WEHI-3CM, augmenting the number of clonable progenitors detected. Optimal activity was observed at 12-25 micrograms protein per plate. Myeloid growth factors (CSF-1 and GM-CSF) were inactive in the murine megakaryocyte assay. The data show lineage specificity for the myeloid stimulators, but a purified erythropoietin preparation was found to stimulate a small level of megakaryocytopoiesis.