Derivation and manipulation of murine embryonic stem cells.
Pluripotent embryonic stem (ES) cell lines were first isolated over 25 years ago and remain an essential tool in molecular and developmental biology to this day. In particular, the use of homologous recombination and subsequent generation of ES-derived mice has greatly facilitated research across all fields. Moreover, ES cells represent an extremely attractive model to study events in early development. In this chapter, we will describe the derivation and propagation of murine ES cells. This is followed by a description of targeting ES cells and a protocol for the generation of mice by diploid and tetraploid blastocyst injections.
Meissner
Alexander
A
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
Eminli
Sarah
S
Jaenisch
Rudolf
R
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation; D002585:Cesarean Section; D002999:Clone Cells; D003470:Culture Media; D004247:DNA / Q000378:metabolism; D004171:Diploidy; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005615:Freezing; D007267:Injections; D051379:Mice; D011123:Polyploidy; D011247:Pregnancy
D003470:Culture Media; D004247:DNA