人急性淋巴细胞白血病细胞株T细胞电转染条件的优化  

Optimization of electransfection for Jurkat T cell

作　　者：江雪[1] 杨平[1,2] 李玥[1] 赵少志[1] 马用信[1] 

机构地区：[1]四川大学华西医院医学遗传室/生物治疗国家重点实验室,成都610041 [2]成都医学院,610041

出　　处：《重庆医学》2011年第15期1463-1465,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(31070676)

摘　　要：目的优化人急性淋巴细胞白血病细胞株细胞(Jurkat T细胞)的电转染条件。方法采用电穿孔的方法,将pEG-FP-C1导入Jurkat T细胞中,通过改变电转时的电压来检测细胞存活率,调节电压、细胞密度、电转质粒数、电转缓冲液,通过流式细胞仪检测质转染效率。结果电转条件在电压140 V、细胞密度107个/毫升、质粒数30μg、电转缓冲液为优化培养基时,电转染效率可达(79.2±3.21)%。本研究为外源基因电转染Jurkat T细胞提供了可靠的试验参数依据。结论本实验通过优化Jur-kat T细胞电转染条件能够有效地提高Jurkat T细胞的电转染效率。Objective To optimize the electransfection parameters of Jurkat T cell and develop an electransfection system for Jurkat T cell.Methods To electransfect the green fluorescent protein（GFP） into Jurkat T cell by electroporation apparatus,with different electric voltages,cell concentrations,plasmid concentration and electroporation buffers.Results The electransfection rat was optimized under the conditions： the electric voltage was 140V,the cell concentration was 107/mL,the plasmid concentration was 30 μg,choosing electroporation buffers as the optimum culture medium.This study provides reliable test parameters for electransfection of Jurkat T cell.Conclusion By optimizing the electransfection conditions,the electransfection rate of Jurkat T cell is efficiently improved.

关 键 词：电转染 急性淋巴细胞白血病细胞株T细胞 转染效率 PEGFP-C1 

分 类 号：R733.7[医药卫生—肿瘤]