title: Efficient iPS cell production with the MyoD transactivation domain in serum-free culture.
abstract: A major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gene composed of Oct4 and the transactivation domain of MyoD (called M(3)O), along with Sox2, Klf4 and c-Myc (SKM). In addition, M(3)O facilitated chromatin remodeling of pluripotency genes in the majority of transduced MEFs, including cells that did not become iPSCs. These observations suggested the possibility that more than 5% of cells had acquired the ability to become iPSCs given more favorable culture conditions. Here, we raised the efficiency of making mouse iPSCs with M(3)O-SKM to 26% by culturing transduced cells at low density in serum-free culture medium. In contrast, the efficiency increased from 0.1% to only 2% with the combination of wild-type Oct4 and SKM (OSKM) under the same culture condition. For human iPSCs, M(3)O-SKM achieved 7% efficiency under a similar serum-free culture condition, in comparison to 1% efficiency with OSKM. This study highlights the power of combining the transactivation domain of MyoD with a favorable culture environment.
authors.affiliation: Stem Cell Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Hirai
authors.forename: Nobuko
authors.initials: N
authors.lastname: Katoku-Kikyo
authors.forename: Peter
authors.initials: P
authors.lastname: Karian
authors.forename: Meri
authors.initials: M
authors.lastname: Firpo
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Kikyo
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D042002:Chromatin Assembly and Disassembly; D018920:Coculture Techniques / Q000379:methods; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence / Q000379:methods; D017570:MyoD Protein / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013724:Teratoma / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: D016895:Culture Media, Serum-Free; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C494174:Pou5f1 protein, mouse; D000469:Alkaline Phosphatase