小鼠B淋巴瘤细胞系A20核转染的影响因素分析  

Factors affecting the transfection efficiency of mouse B lymphoma cell line A20

作　　者：阳诚[1] 宋远 袁小涵 王惠明[1] YANG Cheng;SONG Yuan;YUAN Xiaohan;WANG Huiming(Department of Nephrology,Renmin Hospital of Wuhan University,Wuhan 430060,Chin)

机构地区：[1]武汉大学人民医院肾内科,430060

出　　处：《疑难病杂志》2018年第6期627-629,共3页Chinese Journal of Difficult and Complicated Cases

基　　金：国家自然科学基金资助项目(81370800)

摘　　要：目的分析影响小鼠B淋巴瘤细胞系A20的核转染效率的因素,以提高转染效率。方法2017年1-6月在武汉大学人民医院中心实验室和肾病实验实进行实验。以绿色荧光蛋白(GFP)作为报告基因,使用Nucleofector核酸转染方法分别转染4种外源基因即CD40、ARNO、C-ABL JLP进入A20细胞,用流式细胞术和荧光显微镜分别检测转染效率,研究细胞转染时间和外源质粒大小对核转染效率的影响。结果质粒大小在8 kb以下时转染效率均很高,质粒超过8 kb时转染效率较低,其中,ARNO组(目的基因长度为1 366 bp)转染效率最高,JLP组(目的基因长度为3 976 bp)转染效率最低,转染后4 h检测转染效率最高,48 h时最低。结论核转染方法非常适合于悬浮细胞的转染。且相对于传统的脂质体转染等方式,核转染的方法能有效缩短转染时间。外源基因过大会导致转染效率低,控制外源基因的大小能有效提高转染效率。Objective To investigate the factors affecting the transfection efficiency of mouse B lymphoma cell line A20 and improve transfection efficiency of suspension cells. Methods The B lymphoma A20 cells were transfected with 4 recombinant plasmids containing a green fluorescent protein（ GFP） reporter gene by nucleofection. Then, the transfection efficiency was evaluated by flow cytometry and fluorescence microscopy. The effects of cell transfection time and plasmid size on the efficiency of transfection were assessed. Results The transfection efficiency was high when the plasmid size was below8 KB and the transfection efficiency was highest at 4 hours after transfection. Conclusion Nucleofection is very suitable for transfection of suspension cells with high transfection efficiency. Compared with the lipofection, the nucleofection can effectively shorten the transfection time and improve the transfection efficiency.

关 键 词：B淋巴瘤细胞 A20 核转染 质粒 

分 类 号：R733.1[医药卫生—肿瘤]