title: High-Efficient Transfection of Human Embryonic Stem Cells by Single-Cell Plating and Starvation.
abstract: 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.
authors.affiliation: 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 .
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: 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 .
authors.forename: Caiping
authors.initials: C
authors.lastname: Ren
authors.affiliation: 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 .
authors.forename: Bin
authors.initials: B
authors.lastname: Zhu
authors.affiliation: 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 .
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: 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 .
authors.forename: Weidong
authors.initials: W
authors.lastname: Liu
authors.affiliation: 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 .
authors.forename: Jia
authors.initials: J
authors.lastname: Shi
authors.affiliation: 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 .
authors.forename: Jianxing
authors.initials: J
authors.lastname: Lin
authors.affiliation: 2 Department of Gynecology and Obstetrics, the Second Xiangya Hospital, Central South University , Changsha, People's Republic of China .
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Xia
authors.affiliation: 3 Department of Gynecology and Obstetrics, the Third Xiangya Hospital, Central South University , Changsha, People's Republic of China .
authors.forename: Fei
authors.initials: F
authors.lastname: Zeng
authors.affiliation: 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 .
authors.forename: Jiawen
authors.initials: J
authors.lastname: Chen
authors.affiliation: 4 Department of Neurosurgery, Xiangya Hospital, Central South University , Changsha, People's Republic of China .
authors.forename: Xingjun
authors.initials: X
authors.lastname: Jiang
mesh_terms: D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000235:genetics; D061251:Primary Cell Culture / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering