Skip to main content

Thank you for visiting 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.

Derivation of pluripotent epiblast stem cells from mammalian embryos


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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

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 layers in vitro and in vivo.


  1. 1

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

    ADS  CAS  Article  Google Scholar 

  2. 2

    Martin, G. R. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl Acad. Sci. USA 78, 7634–7638 (1981)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Thomson, J. A. et al. Isolation of a primate embryonic stem cell line. Proc. Natl Acad. Sci. USA 92, 7844–7848 (1995)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Thomson, J. A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147 (1998)

    ADS  CAS  Article  Google Scholar 

  5. 5

    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)

    CAS  Article  Google Scholar 

  6. 6

    Brook, F. A. et al. The derivation of highly germline-competent embryonic stem cells containing NOD-derived genome. Diabetes 52, 205–208 (2003)

    CAS  Article  Google Scholar 

  7. 7

    Vallier, L., Alexander, M. & Pedersen, R. A. Activin/Nodal and FGF pathways cooperate to maintain pluripotency of human embryonic stem cells. J. Cell Sci. 118, 4495–4509 (2005)

    CAS  Article  Google Scholar 

  8. 8

    Pelton, T. A., Sharma, S., Schulz, T. C., Rathjen, J. & Rathjen, P. D. Transient pluripotent cell populations during primitive ectoderm formation: correlation of in vivo and in vitro pluripotent cell development. J. Cell Sci. 115, 329–339 (2002)

    CAS  PubMed  Google Scholar 

  9. 9

    Rathjen, J. et al. Formation of a primitive ectoderm like cell population, EPL cells, from ES cells in response to biologically derived factors. J. Cell Sci. 112, 601–612 (1999)

    CAS  PubMed  Google Scholar 

  10. 10

    Resnick, J. L., Bixler, L. S., Cheng, L. & Donovan, P. J. Long-term proliferation of mouse primordial germ cells in culture. Nature 359, 550–551 (1992)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Matsui, Y., Zsebo, K. & Hogan, B. L. Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture. Cell 70, 841–847 (1992)

    CAS  Article  Google Scholar 

  12. 12

    De Felici, M. & McLaren, A. Isolation of mouse primordial germ cells. Exp. Cell Res. 142, 476–482 (1982)

    CAS  Article  Google Scholar 

  13. 13

    Xu, R. H. et al. BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nature Biotechnol. 20, 1261–1264 (2002)

    CAS  Article  Google Scholar 

  14. 14

    Camus, A., Perea-Gomez, A., Moreau, A. & Collignon, J. Absence of Nodal signaling promotes precocious neural differentiation in the mouse embryo. Dev. Biol. 295, 743–755 (2006)

    CAS  Article  Google Scholar 

  15. 15

    Mesnard, D., Guzman-Ayala, M. & Constam, D. B. Nodal specifies embryonic visceral endoderm and sustains pluripotent cells in the epiblast before overt axial patterning. Development 133, 2497–2505 (2006)

    CAS  PubMed  Google Scholar 

  16. 16

    Nichols, J., Chambers, I., Taga, T. & Smith, A. Physiological rationale for responsiveness of mouse embryonic stem cells to gp130 cytokines. Development 128, 2333–2339 (2001)

    CAS  PubMed  Google Scholar 

  17. 17

    Conlon, F. L. et al. A primary requirement for nodal in the formation and maintenance of the primitive streak in the mouse. Development 120, 1919–1928 (1994)

    CAS  PubMed  Google Scholar 

  18. 18

    Song, J. et al. The type II activin receptors are essential for egg cylinder growth, gastrulation, and rostral head development in mice. Dev. Biol. 213, 157–169 (1999)

    CAS  Article  Google Scholar 

  19. 19

    Gu, Z. et al. The type I serine/threonine kinase receptor ActRIA (ALK2) is required for gastrulation of the mouse embryo. Development 126, 2551–2561 (1999)

    CAS  PubMed  Google Scholar 

  20. 20

    James, D., Noggle, S. A., Swigut, T. & Brivanlou, A. H. Contribution of human embryonic stem cells to mouse blastocysts. Dev. Biol. 295, 90–102 (2006)

    CAS  Article  Google Scholar 

  21. 21

    Boyer, L. A. et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122, 947–956 (2005)

    CAS  Article  Google Scholar 

  22. 22

    Loh, Y. H. et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nature Genet. 38, 431–440 (2006)

    CAS  Article  Google Scholar 

  23. 23

    Rugg-Gunn, P. J., Ferguson-Smith, A. C. & Pedersen, R. A. Epigenetic status of human embryonic stem cells. Nature Genet. 37, 585–587 (2005)

    CAS  Article  Google Scholar 

  24. 24

    Sun, B. W. et al. Temporal and parental-specific expression of imprinted genes in a newly derived Chinese human embryonic stem cell line and embryoid bodies. Hum. Mol. Genet. 15, 65–75 (2006)

    Article  Google Scholar 

  25. 25

    Beddington, R. S. & Robertson, E. J. An assessment of the developmental potential of embryonic stem cells in the midgestation mouse embryo. Development 105, 733–737 (1989)

    CAS  PubMed  PubMed Central  Google Scholar 

  26. 26

    Niwa, H., Miyazaki, J. & Smith, A. G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nature Genet. 24, 372–376 (2000)

    CAS  Article  Google Scholar 

Download references


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 to NOD 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



Corresponding author

Correspondence to Ludovic Vallier.

Ethics declarations

Competing interests

Reprints and permissions information is available at 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)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Brons, I., Smithers, L., Trotter, M. et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448, 191–195 (2007).

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.


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