淋巴细胞不同转染方法比较

目的:用病毒转染和非病毒转染方法将质粒载体转染进淋巴细胞,比较各种方法转染淋巴细胞的效率,确定适合淋巴细胞最佳转染方法。方法:分别用脂质体转染法,电穿孔转染法,慢病毒感染法,核转染法将构建好的酪氨酸羟化酶(TH)基因干扰质粒转染小鼠淋巴细胞,24小时后经荧光显微镜观察增强型绿色荧光蛋白(EGFP)表达水平,确定转染率。结果:核转染法转染TH基因干扰质粒进入淋巴细胞的效率明显高于其他转染和感染方法,24小时后通过观察EGFP的表达确定转染效率可达50%~60%。结论:核转染方法是转染免疫细胞的有效手段,是体外分子生物学方法研究免疫功能的理想手段。

Comparison of different Methods of Transfecting lymphocytes

Objective: Comparing the efficiency of different transfection methods to transfect lymphocytes,with viral and non-viral gene transfection methods to determine the best transfection method.Methods: The plasmid vectors which contained TH genes were transfected with different methods respectively,including Lipofectamine,electroporation,Lenti-viral and nucleofector transfection methods,and the expression levels of enhanced green fluorescent protein(EGFP) 24h after the transfection were calculated to analyses the effect of different transfect conditions with fluorescence microscope.Results: The effectiveness of nucleofector is much higer than by other transfection or infection methods obviously and the effectiveness of transfection can reach 50%~60% by observing the expression level of EGFP.Conclusions: The nucleofector method is an efficient method to transfect immunocytes and is the ideal way to research immunologic function by molecular biology methods in vitro.