Key Laboratory of Animal Reproductive Physiology and Embryo Technology of the Ministry of Agriculture, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi Province, China.
Theriogenology. 2011 Oct 1;76(6):1076-83. doi: 10.1016/j.theriogenology.2011.05.013. Epub 2011 Jul 12.
To improve the efficiency of somatic cell nuclear transfer (SCNT) in goats, we evaluated the effects of the interval between fusion and activation (1 to 5 h), cytochalasin B (CB) treatment after electrofusion, and the number of transferred embryos on the in vivo and in vitro development of cloned caprine embryos. The majority of the reconstructed embryos had condensed chromosomes and metaphase-like chromosomes at 2 and 3 h after fusion; cleavage and blastocyst rates from those two groups were higher (P < 0.05) than those of embryos activated 1, 4, or 5 h after fusion. Treatment with CB between fusion and activation improved in vitro and in vivo development of nuclear transfer (NT) goat embryos by reducing the fragmentation rate (P < 0.05). Although there were no significant differences in NT efficiency, pregnancy rate and kids born per recipient were increased by transfer of 20 or 30 embryos per recipient compared with 10 embryos. We concluded that CB treatment for 2 to 3 h between fusion and activation was an efficient method for generating cloned goats by somatic cell NT. In addition, increasing the number of embryos transferred to each recipient resulted in more live offspring from fewer recipients.
为提高山羊体细胞核移植（SCNT）的效率，我们评估了融合与激活（1 至 5 小时）之间的时间间隔、电融合后细胞松弛素 B（CB）处理以及移植胚胎数量对克隆山羊胚胎体内和体外发育的影响。大多数重构胚胎在融合后 2 至 3 小时具有浓缩的染色体和类似于有丝分裂的染色体；与融合后 1、4 或 5 小时激活的胚胎相比，这两组的卵裂和囊胚形成率更高（P < 0.05）。在融合和激活之间使用 CB 处理可通过降低碎片化率（P < 0.05）来提高核移植（NT）山羊胚胎的体外和体内发育。尽管 NT 效率没有显著差异，但与每受体移植 10 个胚胎相比，每受体移植 20 或 30 个胚胎可增加妊娠率和每个受体出生的幼崽数。我们得出结论，在融合与激活之间进行 2 至 3 小时的 CB 处理是通过体细胞核移植产生克隆山羊的有效方法。此外，增加每个受体移植的胚胎数量可从较少的受体中获得更多的活产后代。
