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Embryonic stem cells require Wnt proteins to prevent differentiation to epiblast stem cells

Nature Cell Biology volume 13, pages 1070–1075 (2011)Cite this article

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Abstract

Pluripotent stem cells exist in naive and primed states, epitomized by mouse embryonic stem cells (ESCs) and the developmentally more advanced epiblast stem cells (EpiSCs; ref. 1). In the naive state of ESCs, the genome has an unusual open conformation and possesses a minimum of repressive epigenetic marks2. In contrast, EpiSCs have activated the epigenetic machinery that supports differentiation towards the embryonic cell types3,4,5,6. The transition from naive to primed pluripotency therefore represents a pivotal event in cellular differentiation. But the signals that control this fundamental differentiation step remain unclear. We show here that paracrine and autocrine Wnt signals are essential self-renewal factors for ESCs, and are required to inhibit their differentiation into EpiSCs. Moreover, we find that Wnt proteins in combination with the cytokine LIF are sufficient to support ESC self-renewal in the absence of any undefined factors, and support the derivation of new ESC lines, including ones from non-permissive mouse strains. Our results not only demonstrate that Wnt signals regulate the naive-to-primed pluripotency transition, but also identify Wnt as an essential and limiting ESC self-renewal factor.

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Figure 1: ESC self-renewal requires Wnt signals.
Figure 2: Wnt signals are required to inhibit the differentiation of ESCs into EpiSCs.
Figure 3: Wnt3a protein is sufficient to inhibit the differentiation of ESCs into EpiSCs.
Figure 4: LIF and Wnt3a are sufficient to support ESC self-renewal.
Figure 5: Wnt3a supports derivation of non-permissive ESCs.

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Change history

  • 07 October 2011

    In the version of this article initially published online, the sequences of the primers used for Wnt2b, Wnt3, Wnt3a, Wnt4, Wnt5a and Wnt5b listed in supplementary table 5 were incorrect.

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Acknowledgements

These studies were supported by the Howard Hughes Medical Institute, the Erasmus MC Stem Cell Institute and grants from the California Institute of Regenerative Medicine (RC1-00133-1), the National Institutes of Health (DK67834-01) and the European Union (FP7-PEOPLE-2009-RG-256560). We thank H. Zeng for technical advice, J. Kong-A-San for assistance with blastocyst injections and R. van der Linden for assistance with FACS. We are grateful for the use of the Cellavista imager and the assistance of V. Vincent and Roche Diagnostics.

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Authors and Affiliations

  1. Department of Cell Biology, Erasmus MC Stem Cell Institute, Erasmus Medical Center, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands

    Derk ten Berge & Dorota Kurek

  2. and Department of Developmental Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA

    Derk ten Berge, Tim Blauwkamp, Wouter Koole, Elif Eroglu, Ronald K. Siu & Roel Nusse

  3. Department of Cell Biology, Erasmus Medical Center, 3000 CA Rotterdam, P.O. Box 2040, The Netherlands

    Alex Maas

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Contributions

D.t.B., D.K. and T.B. designed and carried out experiments, analysed data and wrote the paper. W.K., A.M., R.S. and E.E. designed and carried out experiments and analysed data. R.N. designed experiments and wrote the paper.

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Correspondence to Derk ten Berge.

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ten Berge, D., Kurek, D., Blauwkamp, T. et al. Embryonic stem cells require Wnt proteins to prevent differentiation to epiblast stem cells. Nat Cell Biol 13, 1070–1075 (2011). https://doi.org/10.1038/ncb2314

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