Wang Zhongde
Department of Epigenetics and Embryo Development, Hematech, Inc., Sioux Falls, SD, USA.
Methods Mol Biol. 2011;770:551-69. doi: 10.1007/978-1-61779-210-6_22.
Cloning animals by nuclear transfer (NT) has been successful in several mammalian species. In addition to cloning live animals (reproductive cloning), this technique has also been used in several species to establish cloned embryonic stem (ntES) cell lines from somatic cells. It is the latter application of this technique that has been heralded as being the potential means to produce isogenic embryonic stem cells from patients for cell therapy (therapeutic cloning). These two types of cloning differ only in the steps after cloned embryos are produced: for reproductive cloning the cloned embryos are transferred to surrogate mothers to allow them to develop to full term and for therapeutic cloning the cloned embryos are used to derive ntES cells. In this chapter, a detailed NT protocol in mouse by using somatic stem cells (neuron and skin stem cells) and fully differentiated somatic cells (cumulus cells and fibroblast cells) as nuclear donors is described.
通过核移植（NT）克隆动物在多个哺乳动物物种中已获成功。除了克隆活体动物（生殖性克隆）外，该技术还在多个物种中用于从体细胞建立克隆胚胎干细胞（ntES）系。正是这项技术的后一种应用被誉为从患者身上产生用于细胞治疗的同基因胚胎干细胞的潜在手段（治疗性克隆）。这两种克隆类型仅在克隆胚胎产生后的步骤有所不同：生殖性克隆是将克隆胚胎移植到代孕母体中使其发育至足月，而治疗性克隆则是利用克隆胚胎来获得ntES细胞。在本章中，将详细描述以体干细胞（神经元和皮肤干细胞）以及完全分化的体细胞（卵丘细胞和成纤维细胞）作为核供体的小鼠核移植方案。
