电穿孔转染K562细胞的效率研究

Research of the efficiency of electroporation transfect DNA into K562 cells

摘要: 目的:探讨脂质体转染与电穿孔转染的效率,观察电转染对K562细胞凋亡的影响。方法:以GFP为报告基因,通过脂质体转染及电穿孔两种方式转染K562细胞,荧光显微镜下观察并计算转染效率,Annexin V和PI观察细胞凋亡率。结果:电转染较脂质体转染K562细胞的效率高,电穿孔转染效率10%左右,脂质体转染效率小于1%。电穿孔时K562细胞凋亡率为40%。结论:电转染较脂质体转染K562细胞的效率高,但细胞凋亡率也高。

Abstract: Objective: To obtain higher efficiency in gene transfection into K562 cells and to study the effect of electroporation on cell apoptosis.Methods: Transfection efficiencies with different methods,electroporation and lipofectamine 2000,were calculated under fluorescent microscopy using GFP（Green Fluorescent Protein） as a reporter gene.The frequency of K562 apoptosis was studied with Annexin V and PI.Results: The transfection efficiencies of electroporation（10%）were higher than lipofectamine 2000（＜1%）.The apoptosis frequency of the electroporation transfected K562 cells was 40%.Conclusion: The electroporation is a an ideal method for introduction of foreign gene into the K562 cells.But the electroporation induces cell apoptosis.