Erythroid Differentiation of Murine Embryonic Stem Cells (MESCs) through Modulation of miR-451 and miR-191 Expression

BACKGROUND Expression profile of miRNA in the course of hematopoietic development recommended that miRNAs have potential regulatory roles in hematopoietic differentiation. METHODS In this experimental study, MESCs were infected with lentiviruses containing pCDH-miR-451 and 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-451 expression level increased and miR-191 expression level decreased compared to the untreated MESCs on day 4 post transfection (p< 0.05). Up-regulation of miR-451 and down-regulation of miR-191 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 in these cells (p< 0.001). Moreover, MESCs that were treated with pCDH-miR-451 led to a significant raise of CFU-E colonies compared to untreated control group (p< 0.001). CONCLUSTION Our results showed that up-regulation of miR-451 and down-regulation of miR-191 is essential for erythroid chromatin condensation and strongly promote erythroid differentiation.

pCDH-miR-451, pCDH-anti-miR-191, Murine Embryonic Stem Cells (MESCs), Erythropoiesis