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Induction of pluripotent stem cells from fibroblast cultures

Nature Protocols volume 2, pages 30813089 (2007) | Download Citation

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Abstract

Clinical application of embryonic stem (ES) cells faces difficulties regarding use of embryos, as well as tissue rejection after implantation. One way to circumvent these issues is to generate pluripotent stem cells directly from somatic cells. Somatic cells can be reprogrammed to an embryonic-like state by the injection of a nucleus into an enucleated oocyte or by fusion with ES cells. However, little is known about the mechanisms underlying these processes. We have recently shown that the combination of four transcription factors can generate ES-like pluripotent stem cells directly from mouse fibroblast cultures. The cells, named induced pluripotent stem (iPS) cells, can be differentiated into three germ layers and committed to chimeric mice. Here we describe detailed methods and tips for the generation of iPS cells.

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Acknowledgements

We are grateful to Dr. Yoshimi Tokuzawa for earlier works that were fundamental for our experiments. We also thank Tomoko Ichisaka, Megumi Narita, Yumi Ohuchi, Rie Kato and Ryoko Iyama for technical and administrative supports, and Dr. Toshio Kitamura for pMXs retroviral vectors and Plat-E cells.

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Affiliations

  1. Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-machi, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.

    • Kazutoshi Takahashi
    • , Keisuke Okita
    • , Masato Nakagawa
    •  & Shinya Yamanaka

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Correspondence to Shinya Yamanaka.

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DOI

https://doi.org/10.1038/nprot.2007.418

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