title: Generation of germline-competent rat induced pluripotent stem cells.
abstract: BACKGROUND
  Recent progress in rat pluripotent stem cell technology has been remarkable. Particularly salient is the demonstration that embryonic stem cells (ESCs) in the rat (rESCs) can contribute to germline transmission, permitting generation of gene-modified rats as is now done using mouse ESCs (mESCs) or mouse induced pluripotent stem cells (iPSCs; miPSCs). However, determinations of whether rat iPSCs (riPSCs) can contribute to germ cells are not published. Here we report the germline competency of riPSCs.
  
  
  RESULTS
  We generated riPSCs by transducing three mouse reprogramming factors (Oct3/4, Klf4, and Sox2) into rat somatic cells, followed by culture in the presence of exogenous rat leukemia inhibitory factor (rLIF) and small molecules that specifically inhibit GSK3, MEK, and FGF receptor tyrosine kinases. We found that, like rESCs, our riPSCs can contribute to germline transmission. Furthermore we found, by immunostaining of testis from mouse-rat interspecific chimeras with antibody against mouse vasa homolog, that riPSCs can contribute to embryonic development with chimera formation in mice (rat-mouse interspecific chimeras) and to interspecific germlines.
  
  
  CONCLUSIONS
  Our data clearly demonstrate that using only three reprogramming factors (Oct3/4, Klf4, and Sox2) rat somatic cells can be reprogrammed into a ground state. Our generated riPSCs exhibited germline transmission in either rat-rat intraspecific or mouse-rat interspecific chimeras.
authors.affiliation: Japan Science Technology Agency, Exploratory Research for Advanced Technology, Nakauchi Stem Cell and Organ Regeneration Project, Tokyo, Japan.
authors.forename: Sanae
authors.initials: S
authors.lastname: Hamanaka
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Megumi
authors.initials: M
authors.lastname: Kato-Itoh
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamazaki
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Sato
authors.forename: Ayumi
authors.initials: A
authors.lastname: Umino
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Wakiyama
authors.forename: Mami
authors.initials: M
authors.lastname: Arai
authors.forename: Makoto
authors.initials: M
authors.lastname: Sanbo
authors.forename: Masumi
authors.initials: M
authors.lastname: Hirabayashi
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002678:Chimera / Q000378:metabolism; D046528:Chimerism; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051379:Mice; D051381:Rats; D017208:Rats, Wistar; D013045:Species Specificity