Rhesus monkeys produced by nuclear transfer
Rhesus monkeys produced by nuclear transfer
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DOI：
10.1095/biolreprod57.2.454
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发表时间：
1997-08-01
影响因子：
3.6
通讯作者：
Wolf, DP
中科院分区：
文献类型：
--
作者：
Meng, L;Ely, JJ;Wolf, DP
基因相同的非人灵长类动物可以为基因治疗和研究活动提供强大的动物模型，其中所研究的生理参数直接或间接是可遗传的。在这里，我们证明，核移植是一个可行的技术生产相同的恒河猴。从促性腺激素处理的雌性中回收的卵母细胞通过抽吸第一极体和下面的卵质去核，然后通过放线菌酮暴露激活。单个二倍体卵裂球，回收体外受精产生的胚胎（新鲜或冻融），并用作核供体，下透明的无核（无染色体）卵母细胞和融合的电脉冲注射。重组胚胎在冷冻保存前与布法罗大鼠肝细胞共培养，并转移到同步化的宿主母体。在接受总共29个重组胚胎的9只雌性中，3只怀孕，结果有两个活产，一只雄性和一只雌性。通过7个高度变异的短串联重复基因座的基因型分析，明确确定了两个婴儿的亲子关系。
Genetically identical nonhuman primates can provide a powerful animal model for gene therapy and research activities where the physiological parameters directly or indirectly under study are heritable. Here we demonstrate that nuclear transfer is a viable technology for the production of identical rhesus macaques. Oocytes recovered from gonadotropin-treated females were enucleated by aspiration of the first polar body and underlying ooplasm, then activated by cycloheximide exposure. Individual diploid blastomeres, recovered from in vitro-fertilization-produced embryos (either fresh or frozen-thawed) and used as nuclear donors, were injected under the zona pellucida of enucleated (chromosome-free) oocytes and fused by electric pulses. The reconstituted embryos were cocultured on buffalo rat liver cells before cryostorage and transfer to synchronized host mothers. Of the 9 females receiving a total of 29 reconstituted embryos, 3 became pregnant, with two live births resulting, one male and one female. The parentage of both infants was established unequivocally by genotype analysis at 7 highly variable short tandem repeat loci.