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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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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Takahashi, K., Okita, K., Nakagawa, M. et al. Induction of pluripotent stem cells from fibroblast cultures. Nat Protoc 2, 3081–3089 (2007). https://doi.org/10.1038/nprot.2007.418
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