巨噬细胞RAW264.7不同转染方法的比较

Comparative Study of the Transfection Efficiency of Macrophage by Different Methods

目的:采用不同的转染方法介导siRNA片段转染小鼠巨噬细胞,对不同细胞转染方法进行比较。方法:分别采用脂质体转染、罗氏转染试剂转染、电穿孔法和慢病毒介导siRNA片段转染小鼠巨噬细胞,然后通过流式细胞仪分析不同转染方法的转染效率,并测定对细胞活性的影响以及目的基因的表达水平。结果:慢病毒介导siRNA转染小鼠巨噬细胞效率最高,可达34.75±5.30%,且RNA干扰效果最好;其次为罗氏转染试剂转染和电穿孔法,但电穿孔法细胞活力最低;脂质体转染效率最低,仅为12.17±1.53%,但细胞活力最好。结论:通过对4种小鼠巨噬细胞转染方法的比较,明确了4种转染方法的优缺点,为小鼠巨噬细胞转染提供了实验依据。

Objective： To transfect siRNA into murine macrophage by different methods, and comparison of different transfection methods. Method ：The cells were transfected by different methods with lipofectamine, Roche transfection reagent, electroporation and lentiviral vectors, The analysis of transfection efficiency of different methods by flow cytometer, levels of target mRNA and cell viability were determined. Result： The efficiency of Lentivirus - mediated siRNA transfection is the highest, up to 34. 75 ±5. 30%, and the RNAi effect is best;Followed by Roche transfection reagent and electroporation, but electroporation cell viability lowest;Lipofectamine dif- ficult to transfected, only 12. 17 ± 1.53%, but cell viability of Lipofectamine was greatest. Conclusion： Through the comparison of the four kinds of murine macrophage transfection methods, the advantages and disadvantages of four kinds of transfection methods was ac- quired, and provides the basis for murine macrophage transfection.