Overexpression of microRNA-199b-5p maintains endothelial cells phenotype derived from human induced pluripotent stem cells

Abstract

Objective MicroRNA-199b-5p (miRNA, miR-199b-5p) overexpression was used to determine the effect of miR-199b-5p overexpression on human induced pluripotent stem cells derived endothelial cell (hiPSC-EC) transdifferentiation. Methods Flow cytometry was used to determine endothelial cell markers (CD31+ , CD34+ ) during hiPSC-EC passges at various time points (P1, P5, P10). The expression level of miR-199b-5p was analyzed by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and the endothelial cell markers was measured by flow cytometry at passge 10 after miR-199b-5p overexpression and control lentivirus transduction. Results During passages, hiPSC-EC was detected to have a decreased expression level of CD31and CD34 by flowcytometry, P1 88.69% CD31+ /69.57% CD34+ , P5 68.84% CD31+ /36.90% CD34+ , P10 41.66% CD31+ /19.29% CD34+ and the expression of miR-199b-5p was significantly decreased, hiPSC-ECs overexpressing miR-199b-5p maintained endothelial cell phenotype by demonstrating an increased expression level of CD31 and CD34 as compared to control, miR-199b-5p overexpressing iPSC-ECs P10 74.33% CD31+ /58.56% CD34+ versus control P10 46.30% CD31+ /38.07% CD34+ (F=18.190, P<0.01). Conclusion The expression level of miR-199b-5p was significantly decreased during hiPSC-EC passges and miR-199b-5p overexpression inhibits cell phenotype transition of hiPSCs derived endothelial cells. MiR-199b-5p plays an inhibitory role in iPSC-ECs phenotype transition. Key words: MicroRNA-199b-5p; Induced pluripotent stem cells; Endothelial cells; Phenotype transition