title: Derivation and characterization of mouse embryonic stem cells from permissive and nonpermissive strains.
abstract: Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbred mouse strains. Recently, there have been considerable advances in the field of embryonic stem cell biology, particularly in the area of pluripotency maintenance in the epiblast from which the mESCs are derived. Here we describe a protocol for efficient derivation of germline-competent mESCs from any mouse strain, including strains previously deemed nonpermissive. We provide a protocol that is generally applicable to most inbred strains, as well as a variant for nonpermissive strains. By using this protocol, mESCs can be derived in 3 weeks and fully characterized after an additional 12 weeks, at efficiencies as high as 90% and in any strain background.
authors: Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
authors: Anne
authors: A
authors: Czechanski
authors: Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
authors: Candice
authors: C
authors: Byers
authors: Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
authors: Ian
authors: I
authors: Greenstein
authors: Developmental Biology Program, Sloan-Kettering Institute, New York, New York, USA.
authors: Nadine
authors: N
authors: Schrode
authors: Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
authors: Leah Rae
authors: LR
authors: Donahue
authors: Developmental Biology Program, Sloan-Kettering Institute, New York, New York, USA.
authors: Anna-Katerina
authors: AK
authors: Hadjantonakis
authors: Genetic Resource Science, The Jackson Laboratory, Bar Harbor, Maine, USA.
authors: Laura G
authors: LG
authors: Reinholdt
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line / Q000166:cytology* / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005260:Female; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D011247:Pregnancy