title: A new chemical approach to the efficient generation of mouse embryonic stem cells.
abstract: Here, we present a highly efficient and reproducible method for the establishment of mouse embryonic stem cells (mESCs) from embryonic day 3.5 (E3.5) whole blastocysts. This protocol involves the use of small molecules SB431542 and PD0325901, which inhibit transforming growth factor-β (TGF-β) and extracellular signal-regulated kinases (ERK1/2), respectively. This protocol is universal in the derivation of mESC lines from NMRI, C57BL/6, BALB/c, DBA/2, and FVB/N strains, which have previously been considered refractory or non-permissive for ESC establishment. The efficiency of mESC lines generation is 100%, regardless of genetic background.
authors.affiliation: Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Baharvand
authors.forename: Seyedeh-Nafiseh
authors.initials: SN
authors.lastname: Hassani
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D001755:Blastocyst / Q000166:cytology; D049109:Cell Proliferation; D018920:Coculture Techniques; D015925:Cryopreservation; D003470:Culture Media; D004149:Dioxoles / Q000494:pharmacology*; D004159:Diphenylamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000502:physiology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D061252:Feeder Cells; D051379:Mice; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D003470:Culture Media; D004149:Dioxoles; D018125:Receptors, Transforming Growth Factor beta; C506614:mirdametinib; D004159:Diphenylamine; D048049:Extracellular Signal-Regulated MAP Kinases; D014357:Trypsin