Effects of erythropoietin, IL-3, IL-6 and LIF on a murine megakaryoblastic cell line: Growth enhancement and expression of receptor mRNAs

Abstract

We examined the effects of recombinant human erythropoietin (rhEPO), recombinant murine interleukin 3 (rmIL-3), recombinant human interleukin 6 (rhIL-6), recombinant human interleukin 11 (rhIL-11), recombinant murine leukemia inhibitory factor (rmLIF) and recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) on the growth of murine megakaryocytic cell lines. In serum-free methylcellulose culture supplemented with bovine serum albumin (BSA), the addition of rhEPO (0.12-10 U/ml), rmIL-3 (10-2 500 U/ml), rhIL-6 (100–10000 U/ml), rmLIF (10–1000 U/ml), or rmGM-CSF (10–1000 U/ml) enhanced colony growth in L8057Y5 cells, which had been maintained in protein-free culture, mostly in a dose-dependent fashion; rhIL-11 did not have any stimulatory effect at the tested doses (10–1000 U/ml). In addition, colony growth of L8057 cells, which had been maintained in serum-containing culture, was enhanced, but to a lesser extent, by the addition of these cytokines except rhEPO (the cultures were supplemented with 1% fetal bovine serum. Among the cytokines that showed growth-enhancing effects on L8057 cells, the expression of mRNAs encoding receptors for EPO, IL-6 and IL-3 was examined by northern blot analysis or reverse transcription polymerase chain reaction (RT-PCR). In both cell lines, mRNAs for EPO-R, IL-6R, gp130, IL-3R α and β chains were constitutively expressed. The results suggest that L8057 and L8057Y5 cell lines have characteristics of megakaryoblastic cells in their biological responses to cytokines, as well as in the expression of cytokine receptor mRNAs, and that the growthenhancing effects of these cytokines on the cell lines may be achieved through specific receptors. Our findings show the value of these cell lines for investigating the mechanisms of growth signal transduction in megakaryopoiesis.