title: Generation of Partially Reprogrammed Cells and Fully Reprogrammed iPS Cells by Plasmid Transfection.
abstract: Induced pluripotent stem (iPS) cells can be directly generated from somatic cells by overexpression of defined transcription factors. iPS cells can perpetually self-renew and differentiate into all cell types of an organism. iPS cells were first generated through infection with retroviruses that contain reprogramming factors. However, development of an exogene-free iPS cell generation method is crucial for future therapeutic applications, because integrated exogenes result in the formation of tumors in chimeras and regain pluripotency after differentiation in vitro. Here, we describe a method to generate iPS cells by transfection of plasmid vectors and to convert partially reprogrammed cells into fully reprogrammed iPS cells by switching from mouse ESC culture conditions to KOSR-based media with bFGF. We also describe basic methods used to characterize fully reprogrammed iPS cells.
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Jong Soo
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Hyun Woo
authors.initials: HW
authors.lastname: Choi
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea.
authors.forename: Yean Ju
authors.initials: YJ
authors.lastname: Hong
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, South Korea. dojt@konkuk.ac.kr.|Department of Animal Biotechnology, College of Animal Bioscience and Technology, 120 Neungdong-Ro, Gwangjin-gu, Seoul, 143-701, South Korea. dojt@konkuk.ac.kr.
authors.forename: Jeong Tae
authors.initials: JT
authors.lastname: Do
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000187:drug effects; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D002678:Chimera / Q000235:genetics; D046149:Embryo Culture Techniques / Q000379:methods; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000502:physiology; D008297:Male; D051379:Mice; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000502:physiology; D010957:Plasmids / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000502:physiology; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000502:physiology; D014162:Transfection
chemical_list: D018398:Homeodomain Proteins; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489428:Myc protein, mouse; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D016222:Fibroblast Growth Factor 2
keywords: Conversion; Fully reprogrammed iPS cells; Induced pluripotent stem (iPS) cells; Partially reprogrammed cells; Pluripotency