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Derivation of pluripotent epiblast stem cells from mammalian embryos

Naturevolume 448, pages191–195 (2007)Cite this article

Abstract

Although the first mouse embryonic stem (ES) cell lines were derived 25 years ago1,2 using feeder-layer-based blastocyst cultures, subsequent efforts to extend the approach to other mammals, including both laboratory and domestic species, have been relatively unsuccessful. The most notable exceptions were the derivation of non-human primate ES cell lines3 followed shortly thereafter by their derivation of human ES cells4. Despite the apparent common origin and the similar pluripotency of mouse and human embryonic stem cells, recent studies have revealed that they use different signalling pathways to maintain their pluripotent status. Mouse ES cells depend on leukaemia inhibitory factor and bone morphogenetic protein, whereas their human counterparts rely on activin (INHBA)/nodal (NODAL) and fibroblast growth factor (FGF). Here we show that pluripotent stem cells can be derived from the late epiblast layer of post-implantation mouse and rat embryos using chemically defined, activin-containing culture medium that is sufficient for long-term maintenance of human embryonic stem cells. Our results demonstrate that activin/Nodal signalling has an evolutionarily conserved role in the derivation and the maintenance of pluripotency in these novel stem cells. Epiblast stem cells provide a valuable experimental system for determining whether distinctions between mouse and human embryonic stem cells reflect species differences or diverse temporal origins.

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Figure 1:Derivation of pluripotent epiblast stem cells (EpiSCs) from the late epiblast layer of embryos at post-implantation stages.
Figure 2:Embryonic identity of EpiSCs.
Figure 3:EpiSCs are capable of differentiating into the three primary germ layersin vitro andin vivo.

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Acknowledgements

We thank A. McLaren for her support. We thank A. Smith for CGR8 cells, A. Nagy for R1 cells, P. Andrew for the SSEA-1 antibody, L. Wicker for access toNOD mice, and S. Thiru for advice in the teratoma analysis. This work was supported by an MRC International Appointments Initiative grant (R.A.P), MRC/Juvenile Diabetes Research Foundation Centre funding (R.A.P., I.G.M.B.), Remedi (I.G.M.B.), the Wellcome Trust Functional Genomics Initiative on Stem Cells (L.E.S, M.W.B.T.), the Addenbrookes National Institute for Health Research Biomedical Research Centre, and a Diabetes UK Career Development fellowship (L.V.). We dedicate this paper to the memory of our colleague Isabelle Bouhon.

Author Contributions L.V. conceived the experiment and did the molecular analysis; I.G.M.B. derived and cultured the EpiSCs; R.A.P. performed the epiblast dissections and together with I.G.M.B. carried out the aggregation, chimaera and clonal assays; L.E.S. and M.T. obtained the microarray data; P.R-G. performed the epigenetic analysis; B.S. performed the blastocyst immunosurgery and ICM cultures; S.M.C.d.S.L. did the Stella–GFP dissection and Blimp1 staining; S.K.H. provided PCR analysis of chimaeras; A.C. did karyotyping of human ES cell lines; L.A-R. carried out the teratoma work; L.V., R.A.P. and I.G.M.B. analysed the data and co-wrote the paper.

Author information

Author notes

  1. Peter Rugg-Gunn

    Present address: Present address: Hospital for Sick Children, Toronto Medical Discovery Tower 101 College Street, Toronto, Ontario M5G 1L7, TMDT, Canada.,

Authors and Affiliations

  1. Department of Surgery and Cambridge Institute for Medical Research, Addenbrooke’s Hospital, University of Cambridge, Cambridge CB2 0XY, UK,

    I. Gabrielle M. Brons, Peter Rugg-Gunn, Bowen Sun, Roger A. Pedersen & Ludovic Vallier

  2. CR-UK Viral Oncology Group, Wolfson Institute for Biomedical Research, UCL, Cruciform Building Gower Street, London WC1E 6BT, UK,

    Lucy E. Smithers & Matthew W. B. Trotter

  3. Wellcome Trust/Cancer Research UK Gurdon Institute of Cancer and Developmental Biology and Department of Physiology, University of Cambridge, Tennis Court Road, Cambridge CB2, 1QR, UK,

    Susana M. Chuva de Sousa Lopes

  4. Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, University of Cambridge, Cambridge CB2 0XY, UK,

    Sarah K. Howlett

  5. Medical Genetics Department Cambridge University Hospital NHS Foundation Trust, Kefford House Maris Lane Cambridge CB2 2FF, UK,

    Amanda Clarkson

  6. Dept of Laboratory Medicine Clinical Research Centre, Karolinska University Hospital Karolinska Institutet 141 57 Stockholm, Sweden,

    Lars Ahrlund-Richter

Authors
  1. I. Gabrielle M. Brons
  2. Lucy E. Smithers
  3. Matthew W. B. Trotter
  4. Peter Rugg-Gunn
  5. Bowen Sun
  6. Susana M. Chuva de Sousa Lopes
  7. Sarah K. Howlett
  8. Amanda Clarkson
  9. Lars Ahrlund-Richter
  10. Roger A. Pedersen
  11. Ludovic Vallier

Corresponding author

Correspondence toLudovic Vallier.

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Competing interests

Reprints and permissions information is available atwww.nature.com/reprints. The authors declare no competing financial interests.

Supplementary information

Supplementary Information 1

This file contains Supplementary Data, Supplementary Methods, Supplementary Figures 1-7 with Legends and additional references. (PDF 5757 kb)

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Brons, I., Smithers, L., Trotter, M.et al. Derivation of pluripotent epiblast stem cells from mammalian embryos.Nature448, 191–195 (2007). https://doi.org/10.1038/nature05950

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Editorial Summary

A new type of stem cell

Human embryonic stem (ES) cells are potentially important in therapy because they are pluripotent, capable of differentiating into virtually any cell type given appropriate encouragement. One obstacle to progress in research on them has been the baffling differences between human and mouse ES cells. Now two groups working independently have created a new kind of pluripotent ES cell. Derived from mouse embryos after they implant in the wall of the uterus, these EpiSCs (epiblast stem cells) are distinct from 'classic' mouse ES cells and mirror key features of human ES cells. The discovery of EpiSCs should provide an important experimental model to accelerate the use of human ES cells in research and eventually perhaps, in therapy.

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