THP-1单核细胞不同转染方法的比较

背景：人单核细胞系THP-1细胞是典型的悬浮细胞系,其转染并瞬时表达外源蛋白比一般贴壁细胞相对困难。其中DEAE-葡聚糖介导的细胞转染方法虽然建立较早,但在国内应用很少。目的：采用不同质粒转染方法介导质粒绿色荧光蛋白真核表达质粒（pEGFP-N1）转染人THP-1单核细胞,以获得较高转染效率的方法。方法：分别采用DEAE-葡聚糖、Lipofectamine 2000、FuGENE 6、梭华-Sofast转染试剂不同方法介导质粒绿色荧光蛋白真核表达质粒进行转染,测定不同方法的转染效率、及其对细胞活力的影响。结果与结论：荧光显微镜下观察及流式细胞仪检测结果均显示DEAE-Dextran介导的转染效率最高,Lipfectamine 2000脂质体次之,FuGENE 6和梭华-Sofast则明显低于前二者。将质粒绿色荧光蛋白真核表达质粒体外成功转染人THP-1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。

Comparison of transfection efficiency of THP-1 monocytes by different methods

BACKGROUND： Human THP-1 is a typical suspension cell line. It is more difficult to transfect and transiently express foreign genes into the adherent cells than the suspension cells. Diethylaminoethyl dextran （DEAE-dextran） transfection method has been previously established, but rarely used in China. OBJECTIVE： To transfect plasmid vector coding enhanced green fluorescent protein N1 into human THP-1 monocytes by different methods and to acquire the method with better transfection efficiency. METHODS： The cells were transfected by different methods with DEAE-dextran, Lipofectamine 2000, FuGENE 6 Sofast transfecti0n reagents. Transfection efficiency and cell viability were determined. RESULTS AND CONCLUSION： The transfection efficiency was the highest by DEAE-dextran transfection, then by Lipfectamine 2000, and lowest by FuGENE 6 and Sofast transfection reagent, as detected by fluorescence microscope and flow cytometry. Plasmid enhanced green fluorescent protein N1 was effectively expressed after being transfected into THP-1 monocytes in vitro, and the transfection efficiency was enhanced and the impact on cell viability was reduced by optimizing the transfection methods.