Meissner Alexander, Eminli Sarah, Jaenisch Rudolf
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
Methods Mol Biol. 2009;482:3-19. doi: 10.1007/978-1-59745-060-7_1.
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.
多能胚胎干细胞系于25多年前首次分离出来，至今仍是分子生物学和发育生物学中的重要工具。特别是，同源重组的应用以及随后产生的胚胎干细胞衍生小鼠极大地促进了各个领域的研究。此外，胚胎干细胞是研究早期发育事件极具吸引力的模型。在本章中，我们将描述小鼠胚胎干细胞的分离和培养。接下来将描述胚胎干细胞的靶向以及通过二倍体和四倍体囊胚注射产生小鼠的方案。
