title: Efficient generation of fully reprogrammed human iPS cells via polycistronic retroviral vector and a new cocktail of chemical compounds.
abstract: Direct reprogramming of human somatic cells into induced pluripotent stem (iPS) cells by defined transcription factors (TFs) provides great potential for regenerative medicine and biomedical research. This procedure has many challenges, including low reprogramming efficiency, many partially reprogrammed colonies, somatic coding mutations in the genome, etc. Here, we describe a simple approach for generating fully reprogrammed human iPS cells by using a single polycistronic retroviral vector expressing four human TFs in a single open reading frame (ORF), combined with a cocktail containing three small molecules (Sodium butyrate, SB431542, and PD0325901). Our results demonstrate that human iPS cells generated by this approach express human ES cells markers and exhibit pluripotency demonstrated by their abilities to differentiate into the three germ layers in vitro and in vivo. Notably, this approach not only provides a much faster reprogramming process but also significantly diminishes partially reprogrammed iPS cell colonies, thus facilitating efficient isolation of desired fully reprogrammed iPS cell colonies.
authors: Stem and Progenitor Cell Biology Program, Center for Molecular Medicine, Maine Medical Center Research Institute, Maine Medical Center, Scarborough, Maine, United States of America.
authors: Zhonghui
authors: Z
authors: Zhang
authors: Yongxing
authors: Y
authors: Gao
authors: Albert
authors: A
authors: Gordon
authors: Zack Z
authors: ZZ
authors: Wang
authors: Zhijian
authors: Z
authors: Qian
authors: Wen-Shu
authors: WS
authors: Wu
mesh_terms: D001549:Benzamides; D002087:Butyrates; D018929:Cell Culture Techniques; D065150:Cellular Reprogramming; D004149:Dioxoles; D004159:Diphenylamine / Q000031:analogs & derivatives; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D012190:Retroviridae; D014157:Transcription Factors
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D002087:Butyrates; D004149:Dioxoles; D018125:Receptors, Transforming Growth Factor beta; D014157:Transcription Factors; C506614:mirdametinib; D004159:Diphenylamine