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  • Efficient generation of germ line transmitting chimeras from C57BL/6N ES cells by aggregation with outbred host embryos.

Efficient generation of germ line transmitting chimeras from C57BL/6N ES cells by aggregation with outbred host embryos.

PloS one (2010-06-29)
Marina Gertsenstein, Lauryl M J Nutter, Tammy Reid, Monica Pereira, William L Stanford, Janet Rossant, Andras Nagy
ABSTRACT

Genetically modified mouse strains derived from embryonic stem (ES) cells have become essential tools for functional genomics and biomedical research. Large scale mutagenesis projects are producing libraries of mutant C57BL/6 (B6) ES cells to enable the functional annotation of every gene of the mouse genome. To realize the utility of these resources, efficient and accessible methods of generating mutant mice from these ES cells are necessary. Here, we describe a combination of ICR morula aggregation and a chemically-defined culture medium with widely available and accessible components for the high efficiency generation of germline transmitting chimeras from C57BL/6N ES cells. Together these methods will ease the access of the broader biomedical research community to the publicly available B6 ES cell resources.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
ESGRO Complete Accutase, The ESGRO Complete Accutase is a cell detachment solution of proteolytic & collagenolytic enzymes, qualified for use for the detachment of mouse embryonic stem cells cultured in serum-free conditions with ESGRO Complete Clonal Grade Medium.
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^6 units/ml
Sigma-Aldrich
EmbryoMax® KSOM Mouse Embryo Media, (1X), Liquid, with 1/2 Amino Acids & Phenol Red
Sigma-Aldrich
RESGRO Culture Medium, 250ml, The RESGRO Culture Medium is a specialty media that can be utilized to complement traditional murine Embryonic Stem (ES) cell culture media containing ESGRO murine LIF medium supplement.
Sigma-Aldrich
RESGRO Culture Medium, 500ml, The RESGRO Culture Medium is a complete ready-to-use product that can be utilized to complement traditional murine Embryonic Stem (ES) cell culture media containing ESGRO murine LIF medium supplement. It is available in a 500mL format.