Rational optimization of reprogramming culture conditions for the generation of induced pluripotent stem cells with ultra-high efficiency and fast kinetics

• Published in: Cell research Vol. 21; no. 6; pp. 884 - 894
• Main Authors: Chen, Jiekai, Liu, Jing, Chen, You, Yang, Jiaqi, Chen, Jing, Liu, He, Zhao, Xiangjie, Mo, Kunlun, Song, Hong, Guo, Lin, Chu, Shilong, Wang, Deping, Ding, Ke, Pei, Duanqing
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.06.2011; Nature Publishing Group
• Subjects: 631/532/2064/2158; 631/532/2435; 631/57/2272/1590; Animals; Biomedical and Life Sciences; Cell Biology; Cell culture; Cell Culture Techniques; Cell Dedifferentiation - genetics; Cell fate; Cell Proliferation; Coculture Techniques; Culture Media - chemistry; Culture Media - metabolism; Embryos; Fibroblasts - metabolism; Genetic Vectors; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - physiology; Induced Pluripotent Stem Cells - transplantation; Inhibitory postsynaptic potentials; Karyotyping; Kinetics; KLF4 protein; Kruppel-Like Transcription Factors - genetics; Kruppel-Like Transcription Factors - metabolism; Life Sciences; Mice; Mice, Inbred ICR; Oct-4 protein; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - metabolism; Original; original-article; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Retroviridae - genetics; Somatic cells; SOXB1 Transcription Factors - genetics; SOXB1 Transcription Factors - metabolism; Stem cells; Time Factors; Transcription factors; Transfection; Transplantation Chimera; 优化培养条件; 反应动力学; 多能干细胞; 恢复培养; 胚胎细胞; 诱导; 转录因子; 重新编程; reprogramming; iPSCs; stem cells; culture condition; medium
• ISSN: 1001-0602; 1748-7838
• DOI: 10.1038/cr.2011.51

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 reproarammine by defined factors.