Synergistic effects of erythropoietin and interleukin‐6 on the in vitro proplatelet process formation of rat megakaryocytes

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Abstract
The racombinant hemopolettc factors of megakaryocyte potentlator (MEG-POT) were studied to compare their activity In stimulating proplatelet process formation (PPF) with thrombopoletln (TPO, c-MpI Itgand). For the assay, a highly enriched (>95%) population of more than 90% viable megakaryocytes was Isolated from rat bone marrow using the Immunomagnetic beads method and cultured with fetal calf serum (FCS) or In a serum-free condition. Megakaryocytes developing slender beaded cytoplasmic processes (proplatelet processes) were observed on both inverted phase contract microscopy and scanning electron microscopy. A large number of proplatelet process clusters were dose-dependently formed with the addition of varying doses of recombinant erythropoietin (rEpo) and interleukin-6 (rlL-6) as well as TPO. Epo and IL-6 were demonstrated to act synergistically solely at low doses in the development of PPF (PIn vitro observations, It was suggested that a synergism of Epo and IL-6 might play a significant role in the terminal stage of megakaryocyte maturation leading to platelet release.