title: Establishment of ES cells from inbred strain mice by dual inhibition (2i).
abstract: A number of mouse ES cells from inbred strains have been established to date, but efficiency varies across the different strains. The 129 strain mouse is efficient to establish, whereas C57BL/6 and BALB/c strains are not. It is possible that their genetic backgrounds account for the difference in their ability to establish ES cell lines. In this study, we attempted to establish C57BL/6J and BALB/c Cr ES cells by dual inhibition (2i) using two inhibitors (PD0325901 and CHIR99021) of extracellular signal regulated-kinase (ERK) and glycogen synthase kinase-3 (GSK-3), which promote ES cell differentiation. The results revealed that the establishment efficiencies of C57BL/6J and BALB/c Cr ES cells were remarkably increased by 2i. These ES cells stably expressed pluripotent markers and generated high-contribution chimeras with germline transmission. Furthermore, we generated germline chimeras from C57BL/6J ES cells through the method of gene modification. These findings indicate that 2i is a powerful tool for establishing C57BL/6J and BALB/c Cr ES cells with the ability to generate germline chimeras.
authors: Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima 739-8511, Japan.
authors: Akifumi
authors: A
authors: Kanda
authors: Yusuke
authors: Y
authors: Sotomaru
authors: Seiji
authors: S
authors: Shiozawa
authors: Eiso
authors: E
authors: Hiyama
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002678:Chimera; D004159:Diphenylamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors*; D005260:Female; D005854:Germ Cells / Q000166:cytology*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors*; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*
chemical_list: D001549:Benzamides; C473711:Chir 99021; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011743:Pyrimidines; C506614:mirdametinib; D004159:Diphenylamine; D048049:Extracellular Signal-Regulated MAP Kinases; D038362:Glycogen Synthase Kinase 3