MiRNA-148a promotes the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Studies have shown that miRNA-148a can promote human bone marrow mesenchymal stem cells to differentiate into mature cardiomyocyte-like cells, but the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells has not been reported. OBJECTIVE: To investigate the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells. METHODS: Human induced pluripotent stem cells differentiating into cardiomyocyte-like cells were divided into three groups. Cells in the control group were not treated. Cells in the low expression group were treated with miRNA-148a for 28 days, and those in the high expression group were treated with mimics of miRNA-148a for 28 days. In addition, human induced pluripotent stem cells cultured for 28 days were taken as the blank control group. CCK-8 was used to detect cell proliferation activity. qRT-PCR was used to detect the expression of miRNA-148a. Immunofluorescence staining and western blot analysis were performed to detect the expression of MHC and cTnT protein. RESULTS AND CONCLUSION: The expression of intracellular miR-148a mRNA and cell proliferation activity in the low expression group were lower than those in the blank control and control groups, while those in the high expression group were significantly higher than those in the other three groups (P < 0.01). There were no positive expression of MHC and cTnT in the blank control group. There were positive expressions of MHC and cTnT in the control, low expression and high expression groups. The expression of MHC and cTnT protein in the low expression group was significantly lower than that in the control group, and that in the high expression group was significantly higher than that in the other three groups (P < 0.01). These results suggest that miRNA-148a can promote the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells.