title: Rational optimization of reprogramming culture conditions for the generation of induced pluripotent stem cells with ultra-high efficiency and fast kinetics.
abstract: The ectopic expression of several transcription factors can restore embryonic cell fate to cultured somatic cells and generate induced pluripotent stem cells (iPSCs), revealing a previously unknown pathway to pluripotency. However, this technology is currently limited by low efficiency, slow kinetics and multi-factorial requirement. Here we show that reprogramming can be improved and dramatically accelerated by optimizing culture conditions. First, we developed an optimized defined medium, iCD1, which allows Oct4/Sox2/Klf4 (OSK)-mediated reprogramming to achieve ultra-high efficiency (~10% at day 8). We also found that this optimized condition renders both Sox2 and Klf4 dispensable, although the elimination of these two factors leads to lower efficiency and slower kinetics. Our studies define a shortened route, both in timing and factor requirement, toward pluripotency. This new paradigm not only provides a rationale to further improve iPSC generation but also simplifies the conceptual understanding of reprogramming by defined factors.
authors: Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine at Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors: Jiekai
authors: J
authors: Chen
authors: Jing
authors: J
authors: Liu
authors: You
authors: Y
authors: Chen
authors: Jiaqi
authors: J
authors: Yang
authors: Jing
authors: J
authors: Chen
authors: He
authors: H
authors: Liu
authors: Xiangjie
authors: X
authors: Zhao
authors: Kunlun
authors: K
authors: Mo
authors: Hong
authors: H
authors: Song
authors: Lin
authors: L
authors: Guo
authors: Shilong
authors: S
authors: Chu
authors: Deping
authors: D
authors: Wang
authors: Ke
authors: K
authors: Ding
authors: Duanqing
authors: D
authors: Pei
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D054337:Cell Dedifferentiation / Q000235:genetics*; D049109:Cell Proliferation; D018920:Coculture Techniques; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005822:Genetic Vectors; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology / Q000637:transplantation; D007621:Karyotyping; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D018183:Transplantation Chimera
chemical_list: D003470:Culture Media; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011994:Recombinant Proteins; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse