【摘要】 【目的】雷帕霉素(Rapamycin)可以激活和促进细胞的自噬过程。探明在体外培养系统中添加雷帕霉素,检测是否可以通过促进自噬过程提高iPSCs诱导效率。【方法】在iPSCs诱导过程的1~3d和4~6d分别加入雷帕霉素,检测自噬现象,同时对iPSCs诱导过程中多能性基因表达进行检测,统计原代iPSCs克隆数等,评价雷帕霉素对iPSCs重编程的影响;在iPSCs的培养过程中分别加入50nM/L和100nM/L的雷帕霉素处理。【结果】使用浓度为50nM/L和100nM/L的雷帕霉素处理,可以提高iPSCs的诱导效率。50nM/L浓度处理,重编程效率较高【结论】雷帕霉素在iPSCs重编程过程中可以促进自噬,提高Nanog表达水平,抑制p53表达,进而提高iPSCs诱导效率。更多还原

【Abstract】 【Objective】Rapamycin could activate and accelerate autophagy process of cell.The aim of this experiment was to investigate if rapamycin could increase the inducible efficiency of iPSCs through adding rapamycin in in-vitro cell culture system.【Methods】The autophagy was detected by adding rapamycin to culture system in the day 1-3and day 4-6during the process of iPSCs induction.At the same time,the expression level of pluripotent genes were measured.The amount of primary iPSCs colonies were calculated and the effect of Rapamycin on cell reprogamming was evaluated.50nM/L and 100nM/L rapamycin were used in the process of culture.【Results】The result showed that adding 50nM/L and 100nM/L rapamycin in the culture system could increase the iPSc induction efficiency.And 50nM/L rapamycin demonstrated a better reprogramming efficiency.【Conclusion】In conclusion,rapamycin could improve the inducible efficiency of iPSCs by means of stimulating autophagy,increasing the level of Nanogand inhibiting p53 expressions.更多还原