电穿孔法转染人乳腺癌细胞MCF-7的条件优化研究  

The Optimal Conditions for Efficient Transfection of Human Breast Cancer MCF-7 Cells by Electroporation

作　　者：闫东科 许凤霞 吕平[1] 李冬 Yan Dong-ke;Xu Feng-xia;Lv Ping;Li Dong(College of Biological and Environmental Engineering,Tianjin Vocational Institute,Tianjin 300350)

机构地区：[1]天津职业大学生物与环境工程学院

出　　处：《生物化工》2020年第1期27-31,共5页Biological Chemical Engineering

基　　金：天津市自然科学基金项目（19JCYBJC23500）;天津职业大学科学研究基金项目（No.20181111）

摘　　要：通过控制电穿孔时的电压、电脉冲时长、电脉冲次数、质粒浓度和细胞状态等转染条件,将pcDNA3.1(+)-EGFP质粒转入MCF-7细胞,于倒置荧光显微镜下观察EGFP表达情况以检测转染效率,探讨不同电穿孔条件对人乳腺癌MCF-7细胞转染效率的影响,确定MCF-7细胞的最佳电转染条件。结果表明:人乳腺癌细胞MCF-7在电压150 V、质粒浓度1μg/μL左右、脉冲数2次、脉冲时长5 ms、传代时间24 h时可达到较高的电转染率。By controlling the transfection conditions such as voltage,pulse duration,pulse number,plasmid concentration and cell status,pcDNA3.1(+)-EGFP plasmids were transfected into MCF-7 cells by electroporation under different conditions.Transfection efficiency was detected by inverted fluorescence microscope through observing the expression of EGFP.human breast cancer cells MCF-7 can achieve higher electroporation efficiency via 2 electric pulses at a voltage 150 V,pulse duration of 5 ms,passage time of 24 h,plasmid concentration of 1μg/μL.

关 键 词：电转染 人乳腺癌细胞MCF-7 转染参数 转染效率 

分 类 号：R737.9[医药卫生—肿瘤]