title: From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells.
abstract: Mouse embryonic stem cells (ESCs) derive from the inner cell mass (ICM) of a blastocyst at E3.5 while mouse epiblast stem cells (EpiSCs) derive from the late epiblast of a post-implantation embryo at E5.5-E7.5. Both cells are able to differentiate into derivatives of the three germs layers but only ESCs are able to produce chimeras when they are introduced into a blastocyst. To support the naive state of pluripotency, ESC culture requires Leukemia inhibitory factor (Lif) and either serum or inhibitors of Erk and Gsk3 pathways (2i) while the primed pluripotency of EpiSCs is maintained using Activin A and Fibroblast Growth Factor 2 (FGF2). It is possible to obtain EpiSCs in vitro starting from ESCs but also to induce ESCs starting from EpiSCs even if this second process is very difficult and inefficient. In this protocol we describe how we perform the process of conversion from ESCs to EpiSCs.
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*; D047009:Hathewaya histolytica / Q000201:enzymology; D003470:Culture Media / Q000378:metabolism; D005855:Germ Layers / Q000166:cytology* / Q000378:metabolism; D020784:Matrix Metalloproteinase 8 / Q000378:metabolism; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D003470:Culture Media; D020784:Matrix Metalloproteinase 8
keywords: Chemically defined medium (CDM); Collagenase; Conversion; Epiblast stem cells (EpiSCs); Mouse embryonic stem cells (ESCs)