两种转染人宫颈癌HeLa细胞的方法学比较

目的比较两种不同的转染方法,即Lipofectamine2000和Nucleofector Kit转染人宫颈癌Hela细胞的转染效率。方法分别选取Lipofectamine2000转染和Amax核电转(Amax Nucleofector Kit)两种转染方法。HeLa细胞培养贴壁达到85%~90%后传代获取HeLa单细胞悬液,细胞分成2管,其中1管直接采用核电转法转染红色荧光蛋白(DsRed)后种植于1.5 ml的细胞培养皿;另一管先种植于细胞培养皿,待贴壁24 h后采用Lipofectamine2000转染DsRed。细胞的存活率采用台盼兰进行检测。分别比较两者转染效率及转染前后细胞的存活率,进一步探讨Nucleofector Kit核电转法的高效性。结果 Lipofectamine2000的基因转染率仅为20.23%,而核电转法的转染率为90.14%,后者为前者的4.46倍。Lipofectamine2000转染细胞前后的存活率分别为96.34%和90.76%,而核电转组分别为95.98%和78.35%。结论细胞核电转法能高效稳定地转染人宫颈癌细胞株HeLa细胞。

Comparison of two transfected methods for human cervical carcinoma Hela cells

Objective Human cervical carcinoma cell line HeLa is one of the most common used cells in the present research. In order to get the high transfection,we compared two different transfected methods,which is lipofectamine 2000 and Amax nucleofector kit. Methods We selected two kinds of transfected methods,which were the Lipofectamine2000 transfection and Amax nuclear transfer（Amax nucleofector kit）. The Hela cells were passaged for single cell suspension when the cells adherent culture reached 85% ~90%. The cells were divided into two bottles, and the one was transfected directly with red fluorescent protein（DsRed）,and planted in 1. 5 ml cell culture dish. Another one was transfected the DsRed with lipofectamine2000 after cell plating for 24 h. The transfected efficiency rate,the cell survival rate of the two kinds of methods were compared. Meanwhile,further study were explored about the high efficiency of Nucleofector Kit method. Results The efficiency rate of lipofactamine 2000 was only 20. 23%,however,it can reach up to 90. 14% with nucleofector kit,which was 4. 46 times than the former. The survival rate before and after transfection was 96. 34% and 90. 76%,whereas they were 95. 98% and 78. 35% respectively in nucleofector transfected group. Conclusion The method of using the nucleofector kit can transfected high efficiency for Hela cells.