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

Article metrics


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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


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, public database under accession number GSE13365.


  1. 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)

  2. 2

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

  3. 3

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

  4. 4

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

  5. 5

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

  6. 6

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

  7. 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)

  8. 8

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

  9. 9

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

  10. 10

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

  11. 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)

  12. 12

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

  13. 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)

  14. 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)

  15. 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)

  16. 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)

  17. 17

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

  18. 18

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

  19. 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)

  20. 20

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

  21. 21

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

  22. 22

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

  23. 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)

Download references


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

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) doi:10.1038/nature08235

Download citation

Further reading


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.