Advertisement

Reversion of Mouse Postimplantation Epiblast Stem Cells to a Naïve Pluripotent State by Modulation of Signalling Pathways

  • Astrid Gillich
  • Siqin Bao
  • M. Azim Surani
Part of the Methods in Molecular Biology book series (MIMB, volume 1074)

Abstract

Mouse postimplantation epiblast cultured in activin and basic fibroblast growth factor gives rise to continuously growing epiblast stem cells (EpiSCs) that share key properties with postimplantation epiblast, such as DNA methylation and an inactive X-chromosome. EpiSCs also show a distinct gene expression profile compared to embryonic stem cells (ESCs) derived from preimplantation blastocysts, and do not contribute efficiently to chimeras. EpiSCs can, however, revert to pluripotent ESC-like cells upon exposure to leukemia inhibitory factor–Stat3 signalling on feeder cells. Here we describe a protocol for the establishment of EpiSCs and their reversion to ESCs.

Key words

Epiblast stem cells Embryonic stem cells Postimplantation epiblast Naïve pluripotency Epigenetic reprogramming LIF–Stat3 signalling 

Notes

Acknowledgement

This work was supported by grants from the Wellcome Trust to A.G (RG44593) and to M.A.S.

References

  1. 1.
    Nichols J, Smith A (2009) Naive and primed pluripotent states. Cell Stem Cell 4:487–492PubMedCrossRefGoogle Scholar
  2. 2.
    Borgel J, Guibert S, Li Y et al (2010) Targets and dynamics of promoter DNA methylation during early mouse development. Nat Genet 42:1093–1100PubMedCrossRefGoogle Scholar
  3. 3.
    Monk M, Harper MI (1979) Sequential X chromosome inactivation coupled with cellular differentiation in early mouse embryos. Nature 281:311–313PubMedCrossRefGoogle Scholar
  4. 4.
    Brons IG, Smithers LE, Trotter MW et al (2007) Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448:191–195PubMedCrossRefGoogle Scholar
  5. 5.
    Tesar PJ, Chenoweth JG, Brook FA et al (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448:196–199PubMedCrossRefGoogle Scholar
  6. 6.
    Guo G, Yang J, Nichols J et al (2009) Klf4 reverts developmentally programmed restriction of ground state pluripotency. Development 136:1063–1069PubMedCrossRefGoogle Scholar
  7. 7.
    Bao S, Tang F, Li X et al (2009) Epigenetic reversion of post-implantation epiblast to pluripotent embryonic stem cells. Nature 461:1292–1295PubMedCrossRefGoogle Scholar
  8. 8.
    Yeom YI, Fuhrmann G, Ovitt CE et al (1996) Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells. Development 122:881–894PubMedGoogle Scholar
  9. 9.
    Nagy A (2003) Manipulating the mouse embryo: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  10. 10.
    Hanna J, Markoulaki S, Mitalipova M et al (2009) Metastable pluripotent states in NOD-mouse-derived ESCs. Cell Stem Cell 4:513–524PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Astrid Gillich
    • 1
  • Siqin Bao
    • 1
  • M. Azim Surani
    • 1
  1. 1.Wellcome Trust/Cancer Research UK Gurdon InstituteUniversity of CambridgeCambridgeUK

Personalised recommendations