Gómez Martha C, Pope C Earle
Audubon Nature Center for Research of Endangered Species, 14001 River Road, New Orleans, LA, 70131, USA.
Methods Mol Biol. 2015;1330:133-52. doi: 10.1007/978-1-4939-2848-4_13.
In 2003, the first wild felid was produced by interspecies somatic cell nuclear transfer. Since then other wild felid clone offspring have been produced by using the same technique with minor modifications. This chapter describes detailed protocols used in our laboratory for (1) the isolation, culture, and preparation of fibroblast cells as donor nucleus, and (2) embryo reconstruction with domestic cat enucleated oocytes to produce cloned embryos that develop to the blastocyst stage in vitro and, after transfer into synchronized recipients, establish successful pregnancies.
2003年,首例野生猫科动物通过种间体细胞核移植诞生。自那时起,使用相同技术并稍作修改又培育出了其他野生猫科动物克隆后代。本章详细介绍了我们实验室所使用的方案,内容包括:(1)作为供体细胞核的成纤维细胞的分离、培养和制备;(2)利用家猫去核卵母细胞进行胚胎重构,以产生能在体外发育至囊胚阶段、并在移植到同步受体后成功妊娠的克隆胚胎。