Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Suppression of induced pluripotent stem cell generation by the p53–p21 pathway

Abstract

Induced pluripotent stem (iPS) cells can be generated from somatic cells by the introduction of Oct3/4 (also known as Pou5f1), Sox2, Klf4 and c-Myc, in mouse1,2,3,4 and in human5,6,7,8. The efficiency of this process, however, is low9. Pluripotency can be induced without c-Myc, but with even lower efficiency10,11. A p53 (also known as TP53 in humans and Trp53 in mice) short-interfering RNA (siRNA) was recently shown to promote human iPS cell generation12, but the specificity and mechanisms remain to be determined. Here we report that up to 10% of transduced mouse embryonic fibroblasts lacking p53 became iPS cells, even without the Myc retrovirus. The p53 deletion also promoted the induction of integration-free mouse iPS cells with plasmid transfection. Furthermore, in the p53-null background, iPS cells were generated from terminally differentiated T lymphocytes. The suppression of p53 also increased the efficiency of human iPS cell generation. DNA microarray analyses identified 34 p53-regulated genes that are common in mouse and human fibroblasts. Functional analyses of these genes demonstrate that the p53–p21 pathway serves as a barrier not only in tumorigenicity, but also in iPS cell generation.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: iPS cell generation from p53 -null MEF by three or four reprogramming factors.
Figure 2: T-lymphocyte-derived iPS cells.
Figure 3: p21 as a target of p53 during iPS cell generation.
Figure 4: Effect of p53 suppression on plasmid-mediated mouse iPS cell generation.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data are available at the Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) public database under accession number GSE13365.

References

  1. Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006)

    CAS  Article  Google Scholar 

  2. Maherali, N. et al. Directly reprogrammed fibroblasts show global epigenetic remodelling and widespread tissue contribution. Cell Stem Cell 1, 55–70 (2007)

    CAS  Article  Google Scholar 

  3. Okita, K., Ichisaka, T. & Yamanaka, S. Generation of germ-line competent induced pluripotent stem cells. Nature 448, 313–317 (2007)

    ADS  CAS  Article  Google Scholar 

  4. Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES cell-like state. Nature 448, 318–324 (2007)

    ADS  CAS  Article  Google Scholar 

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

    CAS  Article  Google Scholar 

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

    ADS  CAS  Article  Google Scholar 

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

    ADS  CAS  Article  Google Scholar 

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

    ADS  CAS  Article  Google Scholar 

  9. Yamanaka, S. Strategies and new developments in the generation of patient-specific pluripotent stem cells. Cell Stem Cell 1, 39–49 (2007)

    CAS  Article  Google Scholar 

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

    CAS  Article  Google Scholar 

  11. Wernig, M., Meissner, A., Cassady, J. P. & Jaenisch, R. c-Myc is dispensable for direct reprogramming of mouse fibroblasts. Cell Stem Cell 2, 10–12 (2008)

    CAS  Article  Google Scholar 

  12. Zhao, Y. et al. Two supporting factors greatly improve the efficiency of human iPSC generation. Cell Stem Cell 3, 475–479 (2008)

    CAS  Article  Google Scholar 

  13. de Vries, A. et al. Targeted point mutations of p53 lead to dominant-negative inhibition of wild-type p53 function. Proc. Natl Acad. Sci. USA 99, 2948–2953 (2002)

    ADS  CAS  Article  Google Scholar 

  14. Shinmura, K., Bennett, R. A., Tarapore, P. & Fukasawa, K. Direct evidence for the role of centrosomally localized p53 in the regulation of centrosome duplication. Oncogene 26, 2939–2944 (2007)

    CAS  Article  Google Scholar 

  15. Cecchinelli, B. et al. Ser58 of mouse p53 is the homologue of human Ser46 and is phosphorylated by HIPK2 in apoptosis. Cell Death Differ. 13, 1994–1997 (2006)

    CAS  Article  Google Scholar 

  16. Morita, S., Kojima, T. & Kitamura, T. Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 7, 1063–1066 (2000)

    CAS  Article  Google Scholar 

  17. Bowman, T. et al. Tissue-specific inactivation of p53 tumor suppression in the mouse. Genes Dev. 10, 826–835 (1996)

    CAS  Article  Google Scholar 

  18. Stewart, S. A. et al. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9, 493–501 (2003)

    CAS  Article  Google Scholar 

  19. Okita, K., Nakagawa, M., Hyenjong, H., Ichisaka, T. & Yamanaka, S. Generation of mouse induced pluripotent stem cells without viral vectors. Science 322, 949–953 (2008)

    ADS  CAS  Article  Google Scholar 

  20. Yamanaka, S. Elite and stochastic models for induced pluripotent stem cell generation. Nature 460, 49–52 (2009)

    ADS  CAS  Article  Google Scholar 

  21. Yamanaka, S. A fresh look at iPS cells. Cell 137, 13–17 (2009)

    CAS  Article  Google Scholar 

  22. Aoi, T. et al. Generation of pluripotent stem cells from adult mouse liver and stomach cells. Science 321, 699–702 (2008)

    ADS  CAS  Article  Google Scholar 

  23. Tada, M., Takahama, Y., Abe, K., Nakatsuji, N. & Tada, T. Nuclear reprogramming of somatic cells by in vitro hybridization with ES cells. Curr. Biol. 11, 1553–1558 (2001)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank D. Srivastava for critical reading of the manuscript; M. Narita, A. Okada, N. Takizawa, H. Miyachi and S. Kitano for technical assistance; and R. Kato, S. Takeshima, Y. Ohtsu and E. Nishikawa for administrative assistance. We also thank Y. Sasai and T. Tada for technical advices, T. Kitamura for Plat-E cells and pMXs retroviral vectors, R. Farese for RF8 ES cells, and B. Weinberg and W. Hahn for shRNA constructs. This study was supported in part by a grant from the Leading Project of MEXT, Grants-in-Aid for Scientific Research of JSPS and MEXT, and a grant from the Program for Promotion of Fundamental Studies in Health Sciences of NIBIO (to S.Y.). H. H. is a research student under the Japanese Government (MEXT).

Author Contributions H.H. conducted most of the experiments in this study. K.T. generated iPS cells from T cells and also performed the shRNA experiments. T.I. performed manipulation of mouse embryos, teratoma experiments, and mouse line maintenance. T.A. and O.K. optimized retroviral transduction into T cells. M.N. generated iPS cells with plasmids. K.O. generated the Nanog–GFP reporter mice and the plasmids for iPS cell generation. K.O. and K.T. supervised H.H. S.Y. designed and supervised the study, and prepared the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Shinya Yamanaka.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-3 with Legends and Supplementary Table 1. (PDF 1566 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hong, H., Takahashi, K., Ichisaka, T. et al. Suppression of induced pluripotent stem cell generation by the p53–p21 pathway. Nature 460, 1132–1135 (2009). https://doi.org/10.1038/nature08235

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature08235

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing