应用逆转录病毒载体转染K562细胞株的实验研究  

Experimental study on gene transfer into K562 cells mediated by retroviral vector

作　　者：王存邦[1] 白海[1] 薛志文[1] 杨小亮[1] 

机构地区：[1]兰州军区兰州总医院全军血液病中心,甘肃兰州730050

出　　处：《西北国防医学杂志》2008年第6期405-407,共3页Medical Journal of National Defending Forces in Northwest China

基　　金：全军"十一五"杰出人才基金资助项目(06J005);兰州军区医药科研基金资助项目(LXH-2007006)

摘　　要：目的:探讨多种细胞因子联合刺激下,逆转录病毒载体对K562细胞的基因转移效率。方法:以PA317-GCGPXSN细胞制备病毒上清,NIH3T3细胞测定病毒滴度后,取经过SCF、IL-3、IL-6预培养刺激后的K562细胞,实施基因转移,流式细胞仪及PCR方法测定基因转移效率。结果:①病毒滴度为1.9×105CFU/m l;②对照组K562细胞测定的荧光强度数值为0.56%,实验组为77.16%,两者差异有统计学意义(P<0.01)。PCR方法扩增到了NeoR基因的特异性片断。结论:细胞因子预刺激后实施基因转移,可有效地将外源基因转移进入K562细胞基因组中。Objective:To explore gene transfer efficiency on K562 cells mediated by retroviral vector and stimulated with cytokines.Methods: The viral supernatants were harvested with PA317-GCGPXSN and detected with NIH3T3 cells.Then gene transfer was made using K562 cells as target cells which were stimulated by SCF,IL-3 and IL-6.The gene transfer efficiency was detected by FACS and PCR.Results:The viral titer was 1.9×105 CFU/ml;The gene transfer efficiency of experiment group was 77.16%,whereas the control group was ...

关 键 词：K562细胞 细胞因子 逆转录病毒载体 基因转移 绿色荧光蛋白 

分 类 号：R346[医药卫生—基础医学]