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.

  • Letter
  • Published:

Direct reprogramming of fibroblasts into epiblast stem cells

Nature Cell Biology volume 13, pages 66–71 (2011)Cite this article

Subjects

Abstract

Epiblast stem cells (EpiSCs) derived from epiblast tissue of post-implantation embryos are pluripotent and can give rise to all three germ layers in teratoma assays1,2. Introduction of the four transcription factors Oct4, Sox2, Klf4 and c-Myc into somatic cells has been shown to generate induced pluripotent stem cells (iPSCs) that are very similar to embryonic stem cells (ESCs) in a number of characteristics3,4,5,6. However, generation of EpiSCs by the direct reprogramming of somatic cells using these transcription factors has not been shown to date. Here, we show that these transcription factors can be used to directly generate induced EpiSCs (iEpiSCs) under EpiSC culture conditions. iEpiSCs resemble EpiSCs in morphology, gene expression pattern, epigenetic status and chimaera-forming capability. This study demonstrates that the culture environment in transcription factor-mediated reprogramming determines the cell fate of the reprogrammed cell. We therefore hypothesize that it will eventually be possible to shape the identity of a directly reprogrammed cell simply by modulating culture conditions.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Direct reprogramming of fibroblasts into iEpiSCs.
Figure 2: Characterization of iEpiSCs.
Figure 3: Induction of naive pluripotency in iEpiSCs.
Figure 4: Four factors induce a naive or primed pluripotent state in fibroblasts depending on culture conditions.

Similar content being viewed by others

Accession codes

Accessions

Gene Expression Omnibus

References

  1. Brons, I. G. et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448, 191–195 (2007).

    Article  CAS  Google Scholar 

  2. Tesar, P. J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196–199 (2007).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  5. 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 

  6. 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 

  7. Evans, M. J. & Kaufman, M. H. Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156 (1981).

    Article  CAS  Google Scholar 

  8. Gardner, R. L. & Rossant, J. Investigation of the fate of 4–5 day post-coitum mouse inner cell mass cells by blastocyst injection. J. Embryol. Exp. Morphol. 52, 141–152 (1979).

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  10. Nichols, J. & Smith, A. Naive and primed pluripotent states. Cell Stem Cell 4, 487–492 (2009).

    Article  CAS  Google Scholar 

  11. Rossant, J. Stem cells and early lineage development. Cell 132, 527–531 (2008).

    Article  CAS  Google Scholar 

  12. Guo, G. et al. Klf4 reverts developmentally programmed restriction of ground state pluripotency. Development 136, 1063–1069 (2009).

    Article  CAS  Google Scholar 

  13. Sridharan, R. et al. Role of the murine reprogramming factors in the induction of pluripotency. Cell 136, 364–377 (2009).

    Article  CAS  Google Scholar 

  14. Greber, B. et al. Conserved and divergent roles of FGF signaling in mouse epiblast stem cells and human embryonic stem cells. Cell Stem Cell 6, 215–226 (2010).

    Article  CAS  Google Scholar 

  15. Xu, C. et al. Feeder-free growth of undifferentiated human embryonic stem cells. Nat. Biotechnol. 19, 971–974 (2001).

    Article  CAS  Google Scholar 

  16. Yeom, Y. I. et al. Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells. Development 122, 881–894 (1996).

    CAS  PubMed  Google Scholar 

  17. Bao, S. et al. Epigenetic reversion of post-implantation epiblast to pluripotent embryonic stem cells. Nature 461, 1292–1295 (2009).

    Article  CAS  Google Scholar 

  18. Chou, Y. F. et al. The growth factor environment defines distinct pluripotent ground states in novel blastocyst-derived stem cells. Cell 135, 449–461 (2008).

    Article  CAS  Google Scholar 

  19. Johansson, B. M. & Wiles, M. V. Evidence for involvement of activin A and bone morphogenetic protein 4 in mammalian mesoderm and hematopoietic development. Mol Cell Biol 15, 141–151 (1995).

    Article  CAS  Google Scholar 

  20. Yao, S. et al. Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions. Proc. Natl Acad. Sci. USA 103, 6907–6912 (2006).

    Article  CAS  Google Scholar 

  21. Hadjantonakis, A. K., Gertsenstein, M., Ikawa, M., Okabe, M. & Nagy, A. Non-invasive sexing of preimplantation stage mammalian embryos. Nat. Genet. 19, 220–222 (1998).

    Article  CAS  Google Scholar 

  22. Han, D. W. et al. Epiblast stem cell subpopulations represent mouse embryos of distinct pregastrulation stages. Cell 143, 617–627 (2010).

    Article  CAS  Google Scholar 

  23. Silva, J. et al. Nanog is the gateway to the pluripotent ground state. Cell 138, 722–737 (2009).

    Article  CAS  Google Scholar 

  24. Hayashi, K., Lopes, S. M., Tang, F. & Surani, M. A. Dynamic equilibrium and heterogeneity of mouse pluripotent stem cells with distinct functional and epigenetic states. Cell Stem Cell 3, 391–401 (2008).

    Article  CAS  Google Scholar 

  25. Hanna, J. et al. Metastable pluripotent states in NOD-mouse-derived ESCs. Cell Stem Cell 4, 513–524 (2009).

    Article  CAS  Google Scholar 

  26. Hall, J. et al. Oct4 and LIF/Stat3 additively induce Kruppel factors to sustain embryonic stem cell self-renewal. Cell Stem Cell 5, 597–609 (2009).

    Article  CAS  Google Scholar 

  27. Vierbuchen, T. et al. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463, 1035–1041 (2010).

    Article  CAS  Google Scholar 

  28. Han, D. W. et al. Pluripotential reprogramming of the somatic genome in hybrid cells occurs with the first cell cycle. Stem Cells 26, 445–454 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are indebted to all members of the Schöler laboratory for discussions on the results. We are especially grateful to P. Tesar for providing T9 EpiSCs and A. Malapetsas for editing the manuscript. We are also grateful to M. Sinn for technical assistance and I. M. Verma for providing the pBOB-CAG-iCRE-SD plasmid. This work was supported by funds of the Max Planck Society and the Federal Ministry of Education and Research (BMBF) (Grant 01GN0934).

Author information

Authors and Affiliations

  1. Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, Münster, 48149, Germany

    Dong Wook Han, Boris Greber, Guangming Wu, Natalia Tapia, Marcos J. Araúzo-Bravo, Christof Bernemann, Martin Stehling & Hans R. Schöler

  2. Center for Stem Cell Research, Institute of Biomedical Sciences and Technology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, 143-701, Republic of Korea

    Kinarm Ko

  3. Department of Neuroscience, School of Medicine, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, 143-701, Republic of Korea

    Kinarm Ko

  4. University of Münster, Medical Faculty, Domagkstrasse 3, Münster, 48149, Germany

    Hans R. Schöler

Authors
  1. Dong Wook Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Boris Greber
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guangming Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Natalia Tapia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marcos J. Araúzo-Bravo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kinarm Ko
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christof Bernemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Martin Stehling
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hans R. Schöler
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.W.H. designed and performed most experiments and wrote the manuscript. B.G. performed microarray experiments, analysed the data and edited the manuscript. M.J.A. analysed the microarray data. N.T. carried out lentivirus construction and infection. C.B. analysed the gene expression data. G.W. and K.K. performed morula aggregation and the teratoma assay, respectively. M.S. performed FACS analysis. H.S. conceived the experiments and edited the manuscript.

Corresponding author

Correspondence to Hans R. Schöler.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1288 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Han, D., Greber, B., Wu, G. et al. Direct reprogramming of fibroblasts into epiblast stem cells. Nat Cell Biol 13, 66–71 (2011). https://doi.org/10.1038/ncb2136

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb2136

Search

Advanced 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