Plück Anne, Klasen Christian
Centre for Mouse Genetics, Institute for Genetics, University of Cologne, Koeln, Germany.
Methods Mol Biol. 2009;561:231-43. doi: 10.1007/978-1-60327-019-9_15.
The previous two chapters have described the generation of chimeric embryos by blastocyst microinjection and morula aggregation. This chapter describes the reimplantation of these embryos into pseudopregnant female recipient mice (foster mice) in order for the embryos to develop to term. Four different surgical techniques will be described in detail.The transfer of blastocyst-stage embryos back into the uterus is most commonly used for chimeric embryos generated by microinjection or aggregation, or for rederivation purposes when strains are transferred to a clean environment. Fertilized oocytes manipulated by pronuclear microinjection or two-cell-stage embryos are reintroduced to the infundibulum of the oviduct of pseudopregnant female mice.To generate pseudopregnant females, they need to be mated to sterile male mice. These males can be either generated by vasectomy of stud males or naturally sterile males can be used. Two different techniques for the vasectomy are shown: the vas deferens is accessed through an incision in the scrotal sac or through an abdominal incision.
前两章描述了通过囊胚显微注射和桑椹胚聚集产生嵌合胚胎的方法。本章描述了将这些胚胎重新植入假孕雌性受体小鼠（代孕小鼠）体内，以使胚胎发育至足月的过程。将详细介绍四种不同的手术技术。将囊胚期胚胎移植回子宫最常用于通过显微注射或聚集产生的嵌合胚胎，或者当品系转移到清洁环境时用于重新建立品系的目的。通过原核显微注射操作的受精卵或二细胞期胚胎被重新引入假孕雌性小鼠输卵管的漏斗部。为了产生假孕雌性小鼠，需要将它们与不育雄性小鼠交配。这些雄性小鼠可以通过对种公鼠进行输精管切除术产生，也可以使用自然不育的雄性小鼠。展示了两种不同的输精管切除技术：通过阴囊囊切口或腹部切口进入输精管。
