Abstract:Objective To investigate the effect of miRNA-1 and miRNA-133 overexpression on myocardial differentiation of inducible pluripotent stem cells (iPSC). Methods Embryos from two-week-old American Institute of Cancer Research (ICR) pregnant mice were collected, 0.5 g/L of trypsin was used for digestion of vaccination, mitomycin was used after spreading to three generations, mouse induced pluripotent stem cells (miPSC) was incubated and cultivated at 37℃, miPSC growed on embryonic fibroblasts was digested and passaged. Medium was replaced, miRNA (U6) (10 nmol/L, marked as control group), miRNA-1 (10 nmol/L, marked as miRNA-1 group), miRNA-133 (10 nmol/L, marked as miRNA-133 group), miRNA-1 and miRNA-133 (10 nmol/L respectively, marked as miRNA-1+133 group) were added and transfected for 24 hours, and were washed by PBS, then miPSC differentiation medium was added, and its differentiation was daily observed, cell pulse rate was calculated. RT-PCR was used to detect the expression of miRNA-1, miRNA-133 and cardiac troponin T (cTnT). Results The cTnT was barely expressed in undifferentiated miPSC, compared with 0 day of differentiation, the expreesion level of cTnT of miRNA-1 group and miRNA-133 group was increased after 5 days of differentiation (P < 0.05), the increase was more obvious after 10 days of differentiation (P < 0.01). After 10 days of differentiation, the expression level of cTnT of miRNA-1+133 group was obviously higher than those of miRNA-1 group and miRNA-133 group (P < 0.01). Compared with control group, the cell pulse rate of miRNA-1 group and miRNA-133 group were not increased (P > 0.05), while the cell pulse rate of miRNA-1+133 group was significantly increased (P < 0.01). Conclusion miRNA-1 or miRNA-133 alone does not promote myocardial differentiation, only after the co-expression of miRNA-1 and miRNA-133, it can significantly improve the directional differentiation efficiency of miPSC to cardiac cells.
王芳芳 白宝宝▲ 曾迪 楚轶 李雪. miRNA-1和miRNA-133过表达对诱导性多能干细胞心肌分化的影响[J]. 中国医药导报, 2018, 15(31): 4-7,15.
WANG Fangfang BAI Baobao▲ ZENG Di CHU Yi LI Xue. Effect of miRNA-1 and miRNA-133 overexpression on myocardial differentiation of inducible pluripotent stem cells. 中国医药导报, 2018, 15(31): 4-7,15.