Downregulation of miR-191 Expression Differentiates Murine Embryonic Stem Cells (MESCs) to Erythroid Progenitor Cells

BACKGROUND MiRNAs have been involved in many different cellular processes including metabolism, apoptosis, differentiation, and development. Overexpression of miR-191 blocks erythroid enucleation. Recent studies have demonstrated that down-regulation of miR-191 have a key roles in erythropoiesis. METHODS In this experimental study, MESCs were infected with lentiviruses containing pCDH-anti-miR-191. Erythroid differentiation was estimated using quantitative RT-PCR for transcriptional factors (GATA-1, KLF-1, EPOR) and hemoglobin chains (α, β, γ, ε and ζ) genes and flow cytometery for erythroid surface antigens (TER-119 and CD235a). CFU assay was also performed on days 14th and 21st. RESULTS Mature miR-191 expression level decreased compared to the untreated MESCs on day 4 post transfection (p< 0.001). MiR-191 downregulation could induce transcriptional factors (GATA-1, KLF-1, EPOR) and hemoglobin chains (α, β, γ, ε and ζ) genes expression in MESCs (p< 0.05), and have strong correlation with the appearance of CD235a and Ter-119 markers on these cells (p< 0.05). Moreover, MESCs treated with pCDH-miR-451 led to a significant raise of CFU-E colonies compared to untreated control group (p< 0.05). CONCLUSTION Our results demonstrate that miR-191 down-regulation strongly induces erythroid differentiation and maturation of mESCs. On the other hand, our results provide that down-regulation of miR-191 have the potential role in the production of artificial RBCs without the presence of any stimulatory cytokines.

miR-191, Murine Embryonic Stem Cells (MESCs), Erythropoiesis