体外合成修饰mRNA可转染hUC-MSCs

In vitro synthesized modified mRNA can transfect the hUC-MSCs

目的探讨体外合成修饰的mRNA能否稳定高效的转入人脐带间充质干细胞(hUC-MSC)中,为利用mRNA诱导分化hUC-MSC进行细胞治疗奠定基础。方法体外构建合成mRNA的模板并合成eGFP的修饰mRNA,将其转入hUC-MSCs,流式检测转染效率、最佳转染剂量及mRNA在细胞中的半衰期。相同方法合成hPdx1修饰mRNA转染hUC-MSCs,免疫荧光检测转染效果。结果体外合成的eGFP修饰mRNA可高效转入hUC-MSCs,最佳转染剂量为1.5μg/mL,转染后翻译的蛋白在细胞中可稳定存在3 d。hPdx1的修饰mRNA也可稳定地导入hUC-MSCs。结论体外合成的修饰mRNA稳定性好,可进入细胞并翻译成蛋白。

Objective To investigate the stability and efficiency of the in vitro synthesized modified mRNA, when transfection into human umbilical cord mesenchymal stem cells（hUC-MSCs）. Methods To get the plasmid con- struct as the template for mRNA synthesis and synthesize the mRNA of eGFP. When the modified mRNA was transfected into the hUC-MSCs, using flow cytometry to analyze the transfect efficiency and titrate the best trans- fect dose. The same method was used to synthesize the mRNA of Pdxl and to transfect it into the hUC-MSCs, then to measure the transfect efficiency by immunoflurescence. Results The in vitro synthesized and modified mRNA of eGFP can enter the hUC-MSCs efficiently, the best dose for transfection is 1.5 ~g/mL and the mRNA can stay in the cell stably for 3 days. The in vitro modified mRNA of Pdxl also can enter the hUC-MSCs. Con- clusions In vitro synthesized modified mRNA has good stability and can enter the cells and be translated into protein.