Effect of human recombinant erythropoietin on human marrow megakaryocyte colony formation in vitro

Abstract

The effect of human recombinant erythropoietin (EP) on the growth of human marrow megakaryocyte colony forming units (CFU-M) in vitro was investigated by the use of a plasma clot assay. EP as a single stimulating factor or as an additional factor to optimal concentration of leucocyte conditioned medium (PHA-LCM) had no effect on the number of CFU-M derived colonies. However, addition of EP (0.5-1 U/ml) to cultures with suboptimal concentrations of PHA-LCM increased megakaryocytic colony formation by 50-90% but had no effect on the number of granulocytic-monocytic colonies (CFU-GM). Exposure of marrow cells to EP for 24-48 h in liquid suspension cultures, followed by removal of the hormone and assaying the cells for CFU-M in plasma clots, resulted in a 50-100% increase of megakaryocyte colony formation in vitro. The augmenting effect of EP on CFU-M growth in vitro was abolished when EP was added to the medium after the third day of culture. The presence of factors in human serum and in PHA-LCM was an absolute requirement for the hormone to exert its potentiating effect on human CFU-M growth in vitro. Recombinant EP potentiates the growth of human marrow CFU-M and this effect seems to be exerted during the early stages of CFU-M development in vitro.