Liu Hui, Ren Caiping, Zhu Bin, Wang Lei, Liu Weidong, Shi Jia, Lin Jianxing, Xia Xiaomeng, Zeng Fei, Chen Jiawen, Jiang Xingjun
1 Key Laboratory for Carcinogenesis of Chinese Ministry of Health, Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, School of Basic Medical Science, Central South University , Changsha, People's Republic of China .
2 Department of Gynecology and Obstetrics, the Second Xiangya Hospital, Central South University , Changsha, People's Republic of China .
Stem Cells Dev. 2016 Mar 15;25(6):477-91. doi: 10.1089/scd.2015.0301. Epub 2016 Feb 22.
Nowadays, the low efficiency of small interfering RNA (siRNA) or plasmid DNA (pDNA) transfection is a critical issue in genetic manipulation of human embryonic stem (hES) cells. Development of an efficient transfection method for delivery of siRNAs and plasmids into hES cells becomes more and more imperative. In this study, we tried to modify the traditional transfection protocol by introducing two crucial processes, single-cell plating and starvation, to increase the transfection efficiency in hES cells. Furthermore, we comparatively examined the transfection efficiency of some commercially available siRNA or pDNA transfection reagents in hES cells. Our results showed that the new developed method markedly enhanced the transfection efficiency without influencing the proliferation and pluripotency of hES cells. Lipofectamine RNAiMAX exhibited much higher siRNA transfection efficiency than the other reagents, and FuGENE HD was identified as the best suitable reagent for efficient pDNA transfection of hES cells among the tested reagents.
如今，小干扰RNA（siRNA）或质粒DNA（pDNA）转染效率低下是人类胚胎干细胞（hES细胞）基因操作中的一个关键问题。开发一种将siRNA和质粒高效导入hES细胞的转染方法变得越来越迫切。在本研究中，我们试图通过引入单细胞铺板和饥饿这两个关键步骤来改进传统的转染方案，以提高hES细胞的转染效率。此外，我们比较了一些市售的siRNA或pDNA转染试剂在hES细胞中的转染效率。我们的结果表明，新开发的方法显著提高了转染效率，同时不影响hES细胞的增殖和多能性。Lipofectamine RNAiMAX表现出比其他试剂更高的siRNA转染效率，在测试的试剂中，FuGENE HD被确定为最适合hES细胞高效pDNA转染的试剂。
