Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts (original) (raw)
- Letter
- Published: 30 November 2007
- Michiyo Koyanagi1 na1,
- Koji Tanabe1,
- Kazutoshi Takahashi1,
- Tomoko Ichisaka1,2,
- Takashi Aoi1,
- Keisuke Okita1,
- Yuji Mochiduki1,
- Nanako Takizawa1 &
- …
- Shinya Yamanaka1,2,3,4
Nature Biotechnology volume 26, pages 101–106 (2008)Cite this article
- 21k Accesses
- 2095 Citations
- 66 Altmetric
- Metrics details
Abstract
Direct reprogramming of somatic cells provides an opportunity to generate patient- or disease-specific pluripotent stem cells. Such induced pluripotent stem (iPS) cells were generated from mouse fibroblasts by retroviral transduction of four transcription factors: Oct3/4, Sox2, Klf4 and c-Myc1. Mouse iPS cells are indistinguishable from embryonic stem (ES) cells in many respects and produce germline-competent chimeras2,3,4. Reactivation of the c-Myc retrovirus, however, increases tumorigenicity in the chimeras and progeny mice, hindering clinical applications3. Here we describe a modified protocol for the generation of iPS cells that does not require the Myc retrovirus. With this protocol, we obtained significantly fewer non-iPS background cells, and the iPS cells generated were consistently of high quality. Mice derived from Myc− iPS cells did not develop tumors during the study period. The protocol also enabled efficient isolation of iPS cells without drug selection. Furthermore, we generated human iPS cells from adult dermal fibroblasts without MYC.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Additional access options:
Similar content being viewed by others
References
- Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).
Article CAS Google Scholar - Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES cell-like state. Nature 448, 318–324 (2007).
Article CAS Google Scholar - Okita, K., Ichisaka, T. & Yamanaka, S. Generation of germ-line competent induced pluripotent stem cells. Nature 448, 313–317 (2007).
Article CAS Google Scholar - Maherali, N. et al. Directly reprogrammed fibroblasts show global epigenetic remodelling and widespread tissue contribution. Cell Stem Cell 1, 55–70 (2007).
Article CAS Google Scholar - Chambers, I. et al. Functional expression cloning of nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655 (2003).
Article CAS Google Scholar - Mitsui, K. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642 (2003).
Article CAS Google Scholar - Ryan, A.K. & Rosenfeld, M.G. POU domain family values: flexibility, partnerships, and developmental codes. Genes Dev. 11, 1207–1225 (1997).
Article CAS Google Scholar - Schepers, G.E., Teasdale, R.D. & Koopman, P. Twenty pairs of sox: extent, homology, and nomenclature of the mouse and human sox transcription factor gene families. Dev. Cell 3, 167–170 (2002).
Article CAS Google Scholar - Dang, D.T., Pevsner, J. & Yang, V.W. The biology of the mammalian Kruppel-like family of transcription factors. Int. J. Biochem. Cell Biol. 32, 1103–1121 (2000).
Article CAS Google Scholar - Adhikary, S. & Eilers, M. Transcriptional regulation and transformation by Myc proteins. Nat. Rev. Mol. Cell Biol. 6, 635–645 (2005).
Article CAS Google Scholar - Tokuzawa, Y. et al. Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic stem cell self-renewal and mouse development. Mol. Cell. Biol. 23, 2699–2708 (2003).
Article CAS Google Scholar - Morita, S., Kojima, T. & Kitamura, T. Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 7, 1063–1066 (2000).
Article CAS Google Scholar - Vintersten, K. et al. Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals. Genesis 40, 241–246 (2004).
Article CAS Google Scholar - Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, published online, doi:10.1016/j.cell.2007.11.019 (20 November 2007).
- Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science, published online, doi:10.1126/science.1151526 (20 November 2007).
- Polesskaya, A. et al. Lin-28 binds IGF-2 mRNA and participates in skeletal myogenesis by increasing translation efficiency. Genes Dev. 21, 1125–1138 (2007).
Article CAS Google Scholar - McMahon, A.P. & Bradley, A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell 62, 1073–1085 (1990).
Article CAS Google Scholar - Meiner, V.L. et al. Disruption of the acyl-CoA:cholesterol acyltransferase gene in mice: evidence suggesting multiple cholesterol esterification enzymes in mammals. Proc. Natl. Acad. Sci. USA 93, 14041–14046 (1996).
Article CAS Google Scholar
Acknowledgements
We thank K. Yae and M. Maekawa for their valuable scientific discussions; M. Narita for technical assistance; and R. Kato and R. Iyama for administrative assistance. We also thank K. Tomoda for the RNA of hES cells, T. Kitamura for the Plat-E cells and pMXs retroviral vectors, and R. Farese for the RF8 ES cells. This study was supported in part by a grant from the Program for Promotion of Fundamental Studies in Health Sciences of NIBIO, a grant from the Leading Project of MEXT, a grant from Uehara Memorial Foundation and Grants-in-Aid for Scientific Research of JSPS and MEXT (to S.Y.). T. A. and K.O. are JSPS research fellows.
Author information
Author notes
- Masato Nakagawa and Michiyo Koyanagi: These authors contributed equally to this work.
Authors and Affiliations
- Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 606-8507, Japan
Masato Nakagawa, Michiyo Koyanagi, Koji Tanabe, Kazutoshi Takahashi, Tomoko Ichisaka, Takashi Aoi, Keisuke Okita, Yuji Mochiduki, Nanako Takizawa & Shinya Yamanaka - CREST, Japan Science and Technology Agency, Kawaguchi, 332-0012, Japan
Tomoko Ichisaka & Shinya Yamanaka - Gladstone Institute of Cardiovascular Disease, San Francisco, 94158, CA, USA
Shinya Yamanaka - Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, 606-8507, Japan
Shinya Yamanaka
Authors
- Masato Nakagawa
You can also search for this author inPubMed Google Scholar - Michiyo Koyanagi
You can also search for this author inPubMed Google Scholar - Koji Tanabe
You can also search for this author inPubMed Google Scholar - Kazutoshi Takahashi
You can also search for this author inPubMed Google Scholar - Tomoko Ichisaka
You can also search for this author inPubMed Google Scholar - Takashi Aoi
You can also search for this author inPubMed Google Scholar - Keisuke Okita
You can also search for this author inPubMed Google Scholar - Yuji Mochiduki
You can also search for this author inPubMed Google Scholar - Nanako Takizawa
You can also search for this author inPubMed Google Scholar - Shinya Yamanaka
You can also search for this author inPubMed Google Scholar
Contributions
M.N., M.K., K.Tanabe, K.Takahashi, T.A. and K.O. generated and characterized iPS cells. T.I. performed the chimera experiments. Y.M. prepared plasmids. N.T. characterized iPS cells. S.Y. supervised the study and wrote the manuscript.
Corresponding author
Correspondence toShinya Yamanaka.
Supplementary information
Rights and permissions
About this article
Cite this article
Nakagawa, M., Koyanagi, M., Tanabe, K. et al. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts.Nat Biotechnol 26, 101–106 (2008). https://doi.org/10.1038/nbt1374
- Received: 06 November 2007
- Accepted: 23 November 2007
- Published: 30 November 2007
- Issue Date: January 2008
- DOI: https://doi.org/10.1038/nbt1374