title: A retrospective analysis of germline competence in rat embryonic stem cell lines.
abstract: The factors responsible for conferring germline competence in embryonic stem (ES) cell lines remain unidentified. In the present study, rat ES cell lines (n = 17) were established with 3i medium (SU5402, PD0325901, CHIR99021), 2i medium (PD0325901, CHIR99021) or 2iF medium (PD0325901, CHIR99021, forskolin), and their potential for germline transmission to the G1 generation was examined. Rat strains were divided into an albino group (F344, Wistar or CAG/Venus transgenic rats with the Wistar background) or a colored coat group (Brown-Norway, Dark-Agouti, or BLK rats selected from >F3 generations of Wistar × Dark-Agouti rats based on their black coat color). Successful germline transmission was observed in 57 % (4/7), 40 % (2/5) and 100 % (5/5) of the ES cells established with 3i, 2i and 2iF media, respectively. ES cell lines from the homozygous CAG/Venus transgenic rats were established in all three media, but only the lines established with the 2iF medium were germline-competent. Neither coat-color (albino: 64 %, 7/11; colored: 67 %, 4/6) nor gender of the ES cell lines (XX: 67 %, 2/3; XY: 64 %, 9/14) were likely to affect germline transmission.
authors.affiliation: National Institute for Physiological Sciences, Okazaki, Aichi, Japan. mhirarin@nips.ac.jp
authors.forename: Masumi
authors.initials: M
authors.lastname: Hirabayashi
authors.forename: Chihiro
authors.initials: C
authors.lastname: Tamura
authors.forename: Makoto
authors.initials: M
authors.lastname: Sanbo
authors.forename: Megumi
authors.initials: M
authors.lastname: Kato-Itoh
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hochi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D019070:Cell Lineage / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005854:Germ Cells / Q000166:cytology*; D051381:Rats; D055647:Rats, Transgenic; D012189:Retrospective Studies