Gene targeting and Calcium handling efficiencies in mouse embryonic stem cell lines.

ABSTRACT:

Aim

To compare gene targeting efficiencies, expression profiles, and Ca(2+) handling potentials in two widely used mouse embryonic stem cell lines.

Methods

The two widely used mouse embryonic stem cell lines, R1 and HM-1, were cultured and maintained on Mitomycin C treated mouse embryonic fibroblast feeder cell layers, following standard culture procedures. Cells were incubated with primary and secondary antibodies before fluorescence activated cell sorting analysis to compare known pluripotency markers. Moreover, cells were harvested by trypsinization and transfected with a kinase-inactive murine Tyk2 targeting construct, following the BioRad and Amaxa transfection procedures. Subsequently, the cells were cultured and neomycin-resistant cells were picked after 13 d of selec

SUBMITTER: Mamo S 

PROVIDER: S-EPMC3097933 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature