Electroporation of murine embryonic stem cells: a step-by-step guide.
The manipulation of embryonic stem (ES) cells to generate targeted mutations via homologous recombination has proved an invaluable resource for researchers from fields as diverse as embryology, immunology, physiology, and biophysics. The derivation and culture of these cells has been described in many places, but there is no substitute for hands-on experience. In lieu of that, here we provide a step-by-step guide for the technique of electroporation, complete with micrographs of the cells at each step of the process. This is intended to allow the first time ES cell worker, as well as the more experienced researcher, to view the morphology of ES cells under ideal conditions as well as when the cells are suboptimal and should be discarded.
Tompers
Dennie M
DM
University of Pennsylvania, Department of Cell and Developmental Biology, Philadelphia, Pennsylvania 19104-6058, USA.
Labosky
Patricia A
PA
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D018390:Gene Targeting / Q000379:methods; D051379:Mice; D017410:Practice Guidelines as Topic; D011995:Recombination, Genetic; D013234:Stem Cells / Q000166:cytology*