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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. (Translated from eng). Nature 292(5819):154–156
Martin GR (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. (Translated from eng). Proc Natl Acad Sci U S A 78(12):7634–7638
Brook FA, Gardner RL (1997) The origin and efficient derivation of embryonic stem cells in the mouse. (Translated from eng). Proc Natl Acad Sci U S A 94(11):5709–5712
Smith AG, Heath JK, Donaldson DD et al (1988) Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. (Translated from eng). Nature 336(6200):688–690
Niwa H, Burdon T, Chambers I et al (1998) Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. (Translated from eng). Genes Dev 12(13):2048–2060
Nichols J, Smith A (2009) Naive and primed pluripotent states. Cell Stem Cell 4(6):487–492
Brons IGM, Smithers L, Trotter M et al (2007) Derivation of pluripotent Epiblast Stem Cells from mouse and rat embryos. Nature 448:191–195
Tesar PJ, Chenoweth JG, Brook FA et al (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448:196–199
Wray J, Kalkan T, Smith AG (2010) The ground state of pluripotency. (Translated from eng). Biochem Soc Trans 38(4):1027–1032
Marks H, Kalkan T, Menafra R et al (2012) The transcriptional and epigenomic foundations of ground state pluripotency. (Translated from eng). Cell 149(3):590–604
Habibi E, Brinkman AB, Arand J et al (2013) Whole-genome bisulfite sequencing of two distinct interconvertible DNA methylomes of mouse embryonic stem cells. (Translated from eng). Cell Stem Cell 13(3):360–369
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Tosolini, M., Jouneau, A. (2015). Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 1341. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_207
Download citation
DOI: https://doi.org/10.1007/7651_2015_207
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2953-5
Online ISBN: 978-1-4939-2954-2
eBook Packages: Springer Protocols