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

Naturevolume 460, pages1132–1135 (2009)Cite this article

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. Ap53 (also known asTP53 in humans andTrp53 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 lackingp53 became iPS cells, even without the Myc retrovirus. Thep53 deletion also promoted the induction of integration-free mouse iPS cells with plasmid transfection. Furthermore, in thep53-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.

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Figure 1:iPS cell generation fromp53-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.

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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 numberGSE13365.

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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.

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Authors and Affiliations

  1. Center for iPS Cell Research and Application (CiRA), Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8507, Japan

    Hyenjong Hong, Kazutoshi Takahashi, Tomoko Ichisaka, Takashi Aoi, Masato Nakagawa, Keisuke Okita & Shinya Yamanaka

  2. Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan

    Hyenjong Hong, Masato Nakagawa & Shinya Yamanaka

  3. Yamanaka iPS Cell Special Project, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan

    Tomoko Ichisaka & Shinya Yamanaka

  4. Laboratory for Autoimmune Regulation, RIKEN Center for Allergy and Immunology, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan ,

    Osami Kanagawa

  5. Gladstone Institute of Cardiovascular Disease, San Francisco, California 94158, USA ,

    Shinya Yamanaka

Authors
  1. Hyenjong Hong

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  2. Kazutoshi Takahashi

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  3. Tomoko Ichisaka

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  4. Takashi Aoi

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  5. Osami Kanagawa

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  6. Masato Nakagawa

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  7. Keisuke Okita

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  8. Shinya Yamanaka

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Corresponding author

Correspondence toShinya Yamanaka.

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Supplementary Information

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Hong, H., Takahashi, K., Ichisaka, T.et al. Suppression of induced pluripotent stem cell generation by the p53–p21 pathway.Nature460, 1132–1135 (2009). https://doi.org/10.1038/nature08235

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Editorial Summary

On iPS cells and p53: role of p53–p21 pathway

Induced pluripotent stem (iPS) cells are generated from mouse and human somatic cells by introduction of four genes. Efficiency of this process, however, is low. Here it is reported that up to 10% of transduced mouse embryonic fibroblasts (MEF) lacking p53 became iPS cells, even without the Myc retrovirus. In thep53-null background, iPS cells can be generated from terminally differentiated T lymphocytes. The authors propose that the p53–p21 pathway serves as a barrier not only in tumorigenicity, but also in iPS cell generation.

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