title: Improved generation of C57BL/6J mouse embryonic stem cells in a defined serum-free media.
abstract: C57BL/6 is a well-characterized mouse strain that is used extensively for immunological and neurological research. The establishment of C57BL/6 ES cell lines has facilitated the study of gene-altered mice in a pure genetic background-however, relatively few such lines exist. Using a defined media supplement, knockout serum replacement (KSR) with knockout DMEM (KSR-KDMEM), we find that we can readily establish ES cell lines from blastocysts of C57BL/6J mice. Six lines were established, all of which were karyotypically normal and could be maintained in the undifferentiated state on mouse embryonic fibroblast (MEF) feeders. One line was further tested and found to be karyotypically stable and germline competent, both prior to manipulation and after gene targeting. For this cell line, efficiencies of cell cloning and chimera generation were greater when maintained in KSR-KDMEM. Our work suggests that the use of defined serum-free media may facilitate the generation of ES cells from inbred mouse strains.
authors.affiliation: Genetic Diseases Research Branch, National Human Genome Research Institute, Bethesda, Maryland, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Amalia
authors.initials: A
authors.lastname: Dutra
authors.forename: Aya
authors.initials: A
authors.lastname: Takesono
authors.forename: Lisa
authors.initials: L
authors.lastname: Garrett-Beal
authors.forename: Pamela L
authors.initials: PL
authors.lastname: Schwartzberg
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002460:Cell Line; D002678:Chimera / Q000235:genetics; D016895:Culture Media, Serum-Free; D005260:Female; D018390:Gene Targeting; D005838:Genotype; D007621:Karyotyping; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D013234:Stem Cells / Q000166:cytology*
chemical_list: D016895:Culture Media, Serum-Free