Production of cloned pigs by using somatic cells as donors
Production of cloned pigs by using somatic cells as donors
复制标题
DOI：
10.1089/153623003772032754
复制
发表时间：
2003-12-01
期刊：
影响因子：
--
通讯作者：
Prather, RS
中科院分区：
文献类型：
--
作者：
Lai, LX;Prather, RS
核移植是一个程序，通过它可以创建基因相同的个人。这些核转移技术将应用于农业、生物医学和基础研究。根据供体细胞的来源，核移植可分为胚胎细胞核移植和体细胞核移植。1994年首次在牛中报道了培养细胞的后代（西姆斯和First，1994）。由于这篇论文表明细胞可以在核移植前至少培养28天，因此它为核移植前的细胞遗传修饰带来了巨大的希望，从而有可能产生具有特定遗传修饰的动物。这一概念通过产生转基因绵羊（Schnieke et al.，1997）、猪（Park等人，2001）、小牛（Cibelli等人，1998 a）和基因靶向绵羊（McCreath et al.，2000）和猪（Lai等人，2002 b），源自使用转染体细胞的核移植方法。对于猪，体细胞核移植具有另一个特定的意义，因为它将允许使用遗传修饰程序来从克隆猪产生具有降低的免疫原性的组织和器官以用于异种移植（Lai et al.，2002年b）。然而，体细胞核移植的效率，当作为所用卵母细胞的比例来测量发育到足月时，非常低（1-2%）。通过克隆复制特定基因型的能力受到许多变量的影响，这些变量包括物种、受体卵子的来源、核供体的细胞类型、核移植前供体细胞的处理、人工卵母细胞激活的方法、胚胎培养、重构胚胎核中体细胞印记的可能丢失、移植核的充分重编程失败、以及用于核移植的技术。在猪中，存在额外的困难，因为体外产生的胚胎质量低，并且至少需要四个好的胚胎来启动和建立妊娠。
NUCLEAR TRANSFER is a procedure by which genetically identical individuals can be created. The applications of these techniques for nuclear transfer will be in agricultural, biomedical and basic research. Based on the sources of donor cells, nuclear transfer can be classified into embryonic cell nuclear transfer and somatic cell nuclear transfer. Offspring from cultured cells were first reported in 1994 (Sims and First, 1994) in cattle. Since this paper showed that cells could be cultured at least 28 days prior to nuclear transfer, it held great promise for genetically modifying the cells prior to nuclear transfer, and thus the possibility of producing animals with specific genetic modifications. This concept is exemplified by the generation of transgenic sheep (Schnieke et al., 1997), pigs (Park et al., 2001), calves (Cibelli et al., 1998a), and gene-targeted sheep (McCreath et al., 2000) and pigs (Lai et al., 2002b), derived from nuclear transfer approaches by using transfected somatic cells. For pigs, somatic cell nuclear transfer has another specific significance, as is it would allow the use of genetic modification procedures to produce tissues and organs from cloned pigs with reduced immunogenicity for use in xenotransplantation (Lai et al., 2002b). However, the efficiency of somatic cell nuclear transfer, when measured as development to term as a proportion of oocytes used, has been very low (1–2%). A number of variables influence the ability to reproduce a specific genotype by cloning.These include species, source of recipient ova, cell type of nuclei donor, treatment of donor cells prior to nuclear transfer, the method of artificial oocyte activation, embryo culture, possible loss of somatic imprinting in the nuclei of reconstructed embryos, failure of adequate reprogramming of the transplanted nucleus, and the techniques employed for nuclear transfer. In pig, there are additional difficulties in that the quality of embryos produced in vitro is low, and at least four good embryos are required to initiate and establish a pregnancy.