title: Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium.
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.
authors.affiliation: INRA, UMR1198 Biologie du Développement et Reproduction, F-78350, Jouy-en-Josas, France.
authors.forename: Matteo
authors.initials: M
authors.lastname: Tosolini
authors.affiliation: INRA, UMR1198 Biologie du Développement et Reproduction, F-78350, Jouy-en-Josas, France. alice.jouneau@jouy.inra.fr.
authors.forename: Alice
authors.initials: A
authors.lastname: Jouneau
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D015925:Cryopreservation / Q000379:methods; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000737:chemistry / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D044967:Serum / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003470:Culture Media; D053605:Leukemia Inhibitory Factor; D047428:Protein Kinase Inhibitors; D038362:Glycogen Synthase Kinase 3
keywords: 2i; Chemically defined medium (CDM); Laminin; Mouse embryonic stem cells (ESCs); Serum