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Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium

  • Matteo Tosolini
  • Alice JouneauEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1341)

Abstract

Mouse embryonic stem cells (ESCs) derive from the inner cell mass (ICM) of a blastocyst. These cells are pluripotent and thus able to generate both somatic and germinal lineages. It is possible to maintain ESCs in different pluripotent states depending on the in vitro culture conditions. Classically, ESCs are cultured in the presence of serum and LIF, which sustain the naive state of pluripotency but in this metastable state cells exhibit a large degree of heterogeneity. In the last few years, it has been discovered that when ESCs are cultured in a chemically defined medium (without serum), in the presence of LIF and with the addition of two small molecules (in particular the inhibitors of MAPK and Gsk-3 pathways), they reach a ground state of pluripotency where cells are more homogeneous and more “ICM-like.” In this protocol, we describe how we culture mouse ESCs and the way we switch them from naive to ground state.

Keywords:

Mouse embryonic stem cells (ESCs) Serum 2i Chemically defined medium (CDM) Laminin 

Notes

Acknowledgments

We thank the ANR Programme Investissements d’Avenir REVIVE for their support. MT is a recipient of a PhD fellowship from DIM Biotherapies Ile-de-France.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.INRA, UMR1198 Biologie du Développement et ReproductionJouy-en-JosasFrance

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