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-  2015 

聚乙烯亚胺介导三质粒共转染HEK293细胞的条件优化

DOI: doi:10.7507/1001-5515.20150025

Keywords: 聚乙烯亚胺, 腺相关病毒, HEK293细胞, 转染效率, 条件优化

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Abstract:

本研究用聚乙烯亚胺(PEI)介导表达绿色荧光蛋白(GFP)的rAAV包装系统(pAAV-CMV-GFP、pAAV-RC和pHelper)转染人胚肾293细胞(HEK293细胞)。用ImagePro Plus6.0图像分析软件计算表达GFP的细胞比例来测定1、2、3、4、5、6μg/孔不同质粒剂量, 1∶1、3∶1、5∶1、7∶1不同PEI/质粒比值及转染12、24、36、48、60、72 h不同时间三个变量对转染效率的影响, 以优化PEI介导三质粒共转染HEK293细胞的条件。结果显示, 质粒剂量为4μg/孔、PEI/质粒比值为3∶1~5∶1时转染效率维持较高水平; 转染效率-转染时间曲线为S型增长曲线, 转染效率于60 h达到平台期。优化PEI转染条件能增加三质粒共转染HEK293细胞的转染效率

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