Generation of oligodendrocyte precursor-like cells from mouse embryonic fibroblasts using miR-۷a mimics

Objective(s): Developing effective strategies to enhance remyelination remains a major therapeutic challenge in demyelinating disorders such as multiple sclerosis (MS). MicroRNAs (miRNAs) have emerged as powerful regulators of cell fate. It has been shown that miR-۷a is highly expressed in oligodendrocyte precursor cells (OPCs). This study aimed to investigate whether overexpression of miR-۷a can reprogram mouse embryonic fibroblasts (MEFs) into OPC-like cells and oligodendrocytes in vitro. Materials and Methods: MEFs were cultured, confirmed by vimentin immunostaining, and transfected with miR-۷a mimics using LipofectamineTM ۳۰۰۰ or LipofectamineTM RNAiMAX reagent. Transfection efficiency was assessed by qRT-PCR analysis of miR-۷a expression levels. Cells were then cultured in OPC induction medium and assessed for NG۲ expression at ۱۴ and ۲۱ days post-transfection. Differentiation potential was examined by transferring induced cells into oligodendrocyte differentiation medium and analyzing O۴ and MBP expression using Immunocytofluorescence.Results: LipofectamineTM RNAiMAX-mediated transfection produced significantly higher miR-۷a expression than LipofectamineTM ۳۰۰۰, with a clear dose-dependent effect. MEFs transfected with miR-۷a (۱۰ and ۲۰ pmol) exhibited NG۲-positive cells at both ۱۴ and ۲۱ days, whereas no OPC markers were observed in negative controls or induction medium alone. Furthermore, miR-۷a-induced OPC-like cells differentiated into O۴⁺ and MBP⁺ oligodendrocytes, confirming their lineage potential.Conclusion: Overexpression of miR-۷a successfully reprogrammed MEFs into OPC-like cells and promoted their differentiation into oligodendrocytes in vitro. These findings establish miR-۷a as a potential tool for cell-based reprogramming strategies to generate OPC-like cells, warranting further investigation before translation into in vivo remyelination procedures for MS.

کلیدواژه ها:

Cellular reprogramming techniques ، Fibroblasts ، microRNAs ، MiR-۷a ، Multiple Sclerosis ، Oligodendrocyte precursor cells