Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer.
Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer.
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DOI：
10.1007/978-1-4939-8831-0_6
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Liu C
中科院分区：
文献类型：
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作者：
Du Y;Xie W;Zhang F;Liu C
通过将胚胎干细胞微注射到囊胚中产生嵌合小鼠的能力是传统胚胎干细胞介导敲除技术的关键步骤。近年来，基于设计核酸酶的方法，特别是CRISPR技术，大大减少了对囊胚显微注射的需求。然而，这种方法仍然是一种有价值的技术，用于生成复杂的遗传模型以及干细胞研究。在本章中，我们详细描述了我们实验室使用胚胎干细胞生产嵌合小鼠的过程，包括MEF饲养细胞的衍生和失活，小鼠胚胎干细胞的培养和处理，囊胚的收集和显微注射，最后将注射的囊胚植入假妊娠代孕母亲的子宫。
The ability to generate chimeric mice through microinjecting embryonic stem (ES) cells into blastocysts is a critical step for the conventional ES cell-mediated knockout technology. In recent years, designer nuclease-based methods, especially the CRISPR technology, have substantially decreased the needs for blastocyst microinjection. However, this method has still remained as a valuable technique for generating sophisticated genetic models as well as for stem cell research. In this chapter, we describe the detailed procedures used in our laboratory on how to use ES cells to produce chimeric mice, including derivation and inactivation of MEF feeder cells, culturing and handling of mouse ES cells, collection and microinjection of blastocysts, and finally implantation of injected blastocysts into the uteri of pseudopregnant surrogate mothers.