中文摘要：

目的采用不同质粒转染方法介导重组pIRES2-EGFP-TFPI-2质粒转染U937单核细胞,以获得较高转染效率的方法。方法分别采用电穿孔法、Effectene转染试剂、Lipofectamine 2000转染试剂、电穿孔＋Effectene转染试剂、电穿孔＋Lipofectamine 2000转染试剂及HilyMax转染试剂等不同方法介导质粒进行转染,测定不同方法的转染效率、目的基因mRNA表达水平及其对细胞活力的影响。结果电穿孔＋Effectene转染试剂组、电穿孔＋Lipofectamine 2000组及HilyMax组的转染效率和目的基因mRNA表达较高,且HilyMax组对细胞活力影响较小。结论将重组pIRES2-EGFP-TFPI-2质粒体外成功转染入U937单核细胞中,通过优化转染方法提高转染效率、降低对细胞活力的影响,为基因治疗提供了实验基础。

英文摘要：

Objective To transfect recombinant plasmid pIRES2-EGFP-TFPI-2 into U937 monocytes by different methods,and to acquire the method with better transfection efficiency. Methods The cells were divided into 6 groups transfected by different methods with electroporation,Effectene transfection reagent,Lipofectamine 2000,electroporation＋Effectene transfection reagent,electroporation＋Lipofectamine 2000 and HilyMax transfection reagent.Transfection efficiency,levels of target gene mRNA and cell viability were determined. Results Better transfection efficiency was abtained by electroporation＋Effectene transfection reagent,electroporation＋Lipofectamine 2000 transfection reagent and HilyMax transfection reagent.Moreover,HilyMax transfection reagent had less impaction on cell viability. Conclusions Recombinant plasmid pIRES2-EGFP-TFPI-2 was effectively expressed after being transfected into U937 monocytes in vitro,and the transfection efficiency was enhanced by optimizing the transfection methods,which provides experimental support to further gene therapy.