摘要： 以波尔山羊早期胚胎卵裂球作为核供体进行核移植，比较受体卵母细胞来源以及不同发育阶段胚胎的卵裂球对核移植重构胚的融合率的影响，研究Vero细胞对重构胚体外发育的影响，检测重构胚发育成为个体的能力。结果表明：以体内成熟和体外成熟卵母细胞做核移植受体，重组卵的融合率分别为8.7%和88.7%，二者之间差异不显著(P>0.0)；以来源于8，16，32细胞阶段的卵裂球做核供体，重组卵的融合率分别为91.6%，86%，89.2%，三者之间差异不显著(P>0.0)；融合后的重构胚与Vero细胞单层共培养，卵裂率6.%和桑椹胚率32.7%均高于单纯TCM199培养的3.1%和10.6%(P<0.0)；移植18枚融合后的重构胚于2只受体山羊输卵管，结果未获得妊娠；移植经体内培养得到的3枚桑椹胚于受体山羊子宫内，获得妊娠1只，总体的妊娠率为33%(1/3)，怀孕的受体羊维持妊娠到期，产下1只母羔，初生重3.4 kg，一周岁时体重达到42. kg。通过自然交配产下一对波尔山羊羔。

关键词: 山羊, 胚胎细胞, 核移植, 电融合, 克隆, 波尔山羊

Abstract: The aim of the present study was to carry out nuclear transfer with the Boer goat embryo blastomeres as nuclear donors, to compare the effects of the sources of recipient oocytes and development stages of donor embryos on the fusion rate of reconstructed embryo; to investigate if Vero cell monolayers improved the development of reconstructed embryo in vitro; the reconstructed embryos developed to term. The fusion rate of reconstructed embryos from in vivo or in vitro matured oocytes was 85.7% and 88.7% respectively, there was no significant difference between them (P>0.05). The fusion rate of reconstructed embryos with blastomeres from 8, 16, 32 cell embryos was 91.6%, 86%, 89.2%, there was no significant difference among them (P>0.05). Co-cultured with Vero cell monolayers, the reconstructed embryos obtained higher clevaged and morula rate (65.5%, 32.7%) than that of simply cultured in TCM199 (53.1%, 10.6%)(P<0.05). Eighteen fused reconstructed embryos were transferred into 2 recipients, no pregnancy was confirmed; three morula cultured in vivo were transferred into 1 recipient, the recipient was confirmed pregnant (33%), the pregnance was maintained through term, and gave birth a female kid. The kid weighed 3.45 kg, and grew to 42.5 kg one years old. The cloned goat gave birth a twin Boer kids by natural mating, this is the first report of cloned Boer goat produced by embryo cell nuclear transfer.

Key words: Goat, Embronic cells, Nuclear transfer, Electric fusion, Cloning, Boer goat

中图分类号: 

• S827.9+2