淋巴细胞不同转染方法比较

Comparison of different Methods of Transfecting lymphocytes

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。

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.