THP-1单核细胞不同转染方法的比较  

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

作　　者：葛晶[1] 成蓓[1] 彭雯[1] 李云桥[1] 翟伟[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院老年病科,湖北省武汉市430022

出　　处：《中国组织工程研究》2014年第38期6105-6109,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30471921);湖北省自然科学基金面上项目(2013CFB073)~~

摘　　要：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。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.

关 键 词：单核细胞 转染 质粒 组织构建 组织工程 转染效率 THP-1单核细胞 DEAE-葡聚糖 脂质体 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]