Pan Xiaoyan, Zhang Yanli, Guo Zhiqin, Wang Feng
Cell Biol Int. 2014 Feb;38(2):211-8. doi: 10.1002/cbin.10191.
Interspecies nuclear transfer has already achieved success in several species, which shows great potential in recovery and conservation of endangered animals. The study was conducted to establish an efficient system for in vitro argali (Ovis ammon)-sheep embryo reconstruction via interspecies somatic cell nuclear transfer (iSCNT). The competence of domestic sheep cytoplasts to reprogram the adult argali fibroblast nuclei was evaluated, and the effects of enucleation methods and donor cell passage and cell state on the in vitro development of argali-sheep cloned embryos were also examined. Sheep oocytes could support argali and sheep fibroblast cell nuclei transfer and develop to blastocysts in vitro. Oocytes matured for 21–23 h and enucleated by chemically assisted enucleation (CAE) had a higher enucleation rate than blind enucleation (BE), but the development rate of iSCNTembryos was the same (P>0.05). Moreover, passage numbers of fibroblast cells <10, as well as the cell cycle stages did not affect the development rate of iSCNT reconstructed embryos. Thus sheep cytoplasm successfully supports argali nucleus development to blastocyst stage after optimising the nuclear transfer procedure, which indicates that iSCNT can be used to conserve endangered argali in the near future.
种间核移植已在多个物种中取得成功，这在濒危动物的恢复和保护方面显示出巨大潜力。本研究旨在通过种间体细胞核移植（iSCNT）建立一种高效的盘羊（Ovis ammon）-绵羊体外胚胎重建系统。评估了家羊细胞质对成年盘羊成纤维细胞核重编程的能力，并研究了去核方法、供体细胞传代次数和细胞状态对盘羊-绵羊克隆胚胎体外发育的影响。绵羊卵母细胞能够支持盘羊和绵羊成纤维细胞核移植，并在体外发育至囊胚阶段。成熟21-23小时并通过化学辅助去核（CAE）去核的卵母细胞去核率高于盲吸去核（BE），但iSCNT胚胎的发育率相同（P>0.05）。此外，成纤维细胞传代次数<10以及细胞周期阶段均不影响iSCNT重构胚胎的发育率。因此，在优化核移植程序后，绵羊细胞质成功支持盘羊细胞核发育至囊胚阶段，这表明iSCNT在不久的将来可用于保护濒危盘羊。
