三种阳离子转染试剂对小鼠血管瘤内皮细胞转染效率及细胞毒性的影响

目的 从三种常用的阳离子转染试剂中筛选出对小鼠血管瘤内皮细胞(EOMA)有较高转染效率和较低细胞毒性的转染试剂.方法 以含有增强型绿色荧光蛋白(EGFP)报告基因的真核表达载体pEGFP-N1为报告基因,以阳离子脂质体LipofectamineTM2000、LipofectamineTM PLUS和阳离子聚合物JetPEITM为转染试剂,按照转染试剂盒的说明优化其转染条件,分别转染EOMA细胞,24 h后在荧光显微镜下计数阳性细胞率、MTT法检测各转染条件下对EOMA的细胞毒性.结果 经优化转染条件,LipofectamineTM PLUS和JetPEITM转染的细胞中阳性细胞的最高比例分别为45%和47%,LipofectamineTM2000转染的细胞中荧光蛋白表达的比例最高(＞80%),Lipofecta-mineTM2000试剂在最高转染效率的剂量下仍然保证了80%的细胞存活率.结论 阳离子脂质体LipofectamineTM2000对小鼠EOMA细胞有较高的转染效率和较低的细胞毒作用.

Effects of three cationic transfection reagents on transfection efficiency and cytotoxicity of EOMA Cell

Objective To filtrate the transfection reagent with high transfection efficiency and low cytotoxicity to EOMA cell from three cationic transfection reagents. Methods pEGFP-N1 plasmid as a report gene, cationic liposome LipofectamineTM 2000, LipofectamineTM PLUS and cationic polymer JetPEITM were used as the transfection reagents. The positive cells were counted by fluorescence microscopy and the cell viability was calculated by MTT assay in different transfection conditions. Results By optimizing the transfection condition, the highest transfection efficiency of LipofectamineTM PLUS or JetPEI^TM was 45% or 47%, and the positive cell ratio of LipofectamineTM 2000 was over 80 % with about 80 % cell livability. Conclusion LipofectamineTM 2000 is a safe and effective transfection reagent to EOMA cells.