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Generation of human-induced pluripotent stem cells

Nature Protocols volume 3, pages 1180–1186 (2008)Cite this article

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Abstract

Pluripotent cells, such as embryonic stem cells, are invaluable tools for research and can potentially serve as a source of cell- and tissue-replacement therapy. Rejection after transplantation of cells and tissue derived from embryonic stem cells is a significant obstacle to their clinical use. Recently, human somatic cells have been reprogrammed directly to pluripotency by ectopic expression of four transcription factors (Oct4, Sox2, Klf4 and Myc) to yield induced pluripotent stem (iPS) cells. Human iPS cells are a potential source of patient-specific pluripotent stem cells that would bypass immune rejection. iPS cells can also be used to study diseases for which there are no adequate human in vitro or animal models. In this protocol, we describe how to establish primary human fibroblasts lines and how to derive iPS cells by retroviral transduction of reprogramming factors. Overall, it takes 2 months to complete reprogramming human primary fibroblasts starting from biopsy.

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Figure 1: Overview of preparing human dermal fibroblasts and isolating induced pluripotent stem (iPS) cells.
Figure 2: Induced pluripotent stem (iPS) cells have characteristics resembling those of human embryonic stem (hES) cells.

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Change history

  • 04 September 2008

    In Figure 2a of the PDF version of this article initially published online, five labels (OCT4, SOX2, MYC, KLF4 and βACT) were omitted from the left side of the figure. The error has been corrected in the PDF version of the article.

References

  1. Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007).

    Article  CAS  PubMed  Google Scholar 

  2. Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007).

    Article  CAS  PubMed  Google Scholar 

  3. Park, I.H. et al. Reprogramming of human somatic cells to pluripotency with defined factors. Nature 451, 141–146 (2008).

    Article  CAS  PubMed  Google Scholar 

  4. Nakagawa, M. et al. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat. Biotechnol. 26, 101–106 (2008).

    Article  CAS  PubMed  Google Scholar 

  5. Lowry, W.E. et al. Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc. Natl. Acad. Sci. USA 105, 2883–2888 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Takahashi, K., Okita, K., Nakagawa, M. & Yamanaka, S. Induction of pluripotent stem cells from fibroblast cultures. Nat. Protoc. 2, 3081–3089 (2007).

    Article  CAS  PubMed  Google Scholar 

  7. Tiscornia, G., Singer, O. & Verma, I.M. Production and purification of lentiviral vectors. Nat. Protoc. 1, 241–245 (2006).

    Article  CAS  PubMed  Google Scholar 

  8. Lerou, P.H. et al. Derivation and maintenance of human embryonic stem cells. Nat. Protoc. 3, 923–933 (2008).

    Article  CAS  PubMed  Google Scholar 

  9. Tian, X. & Kaufman, D.S. Hematopoietic development of human embryonic stem cells in culture. Methods Mol. Biol. 290, 149–162 (2005).

    PubMed  Google Scholar 

Download references

Acknowledgements

This work was made possible through the generosity and vision of Jonathan and Patti Kraft and private funds donated to the Children's Hospital Boston. G.Q.D. is supported by grants from the National Institutes of Health and the NIH Director's Pioneer Award, and the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research.

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Author notes

  1. In-Hyun Park and Paul H Lerou: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Pediatric Hematology Oncology, Children's Hospital Boston, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    In-Hyun Park, Rui Zhao, Hongguang Huo & George Q Daley

  2. Division of Newborn Medicine, Brigham and Women's Hospital and Children's Hospital Boston, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Paul H Lerou

  3. Division of Hematology, Brigham and Women's Hospital, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    George Q Daley

  4. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    George Q Daley

  5. Harvard Stem Cell Institute, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA

    George Q Daley

  6. Howard Hughes Medical Institute, Boston, 02115, Massachusetts, USA

    George Q Daley

Authors
  1. In-Hyun Park
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  2. Paul H Lerou
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  3. Rui Zhao
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  4. Hongguang Huo
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  5. George Q Daley
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Corresponding author

Correspondence to George Q Daley.

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Park, IH., Lerou, P., Zhao, R. et al. Generation of human-induced pluripotent stem cells. Nat Protoc 3, 1180–1186 (2008). https://doi.org/10.1038/nprot.2008.92

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