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Improved generation of C57BL/6J mouse embryonic stem cells in a defined serum‐free media
Author(s) -
Cheng Jun,
Dutra Amalia,
Takesono Aya,
GarrettBeal Lisa,
Schwartzberg Pamela L.
Publication year - 2004
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.20031
Subject(s) - chimera (genetics) , embryonic stem cell , biology , germline , cell culture , inbred strain , mouse strain , cloning (programming) , microbiology and biotechnology , c57bl/6 , knockout mouse , homologous recombination , gene , genetics , gene knockout , stem cell , immunology , computer science , programming language
Abstract Summary: 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. genesis 39:100–104, 2004. Published 2004 Wiley‐Liss, Inc.