Protocol abstract


Nature Protocols 3, 1180 - 1186 (2008)
Published online: 26 June 2008 | Corrected online: 4 September 2008 | doi:10.1038/nprot.2008.92

Subject Categories: Cell and tissue culture | Isolation, purification and separation

Generation of human-induced pluripotent stem cells

In-Hyun Park1,7, Paul H Lerou2,7, Rui Zhao1, Hongguang Huo1 & George Q Daley1,3,4,5,6


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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  1. Division of Pediatric Hematology Oncology, Children's Hospital Boston, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.
  2. Division of Newborn Medicine, Brigham and Women's Hospital and Children's Hospital Boston, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.
  3. Division of Hematology, Brigham and Women's Hospital, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.
  4. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.
  5. Harvard Stem Cell Institute, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.
  6. Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA.
  7. These authors contributed equally to this work.

Correspondence to: George Q Daley1,3,4,5,6 e-mail: george.daley@childrens.harvard.edu