【作者】 雷安民； 敬晓棋； 严兴荣； 胥涛； 窦忠英；

【Author】 LEI An-min, JING Xiao-qi, YAN Xing-rong, XU Tao, DOU Zhong-ying(Shaanxi Sub-Center of National Stem Cell Engineering &Technology, Northwest Sci-Tech University of Agriculture &Forestry, Yangling, Shaanxi 712100, China)

【机构】 西北农林科技大学陕西省干细胞工程技术研究中心；

【摘要】 应用相同体外成熟培养液（TCM199液内添加0.22mg/L丙酮酸钠,10nm Hepes,0.10IU HMG,1 mg/mL17B-E₂,10%胎牛血清）对牛、羊卵泡卵母细胞进行体外成熟培养,然后挑选具有第一极体的成熟卵母细胞,去核后分别与山羊耳上皮细胞构建克隆胚胎。挑选成功融合的同种（羊-羊）克隆胚胎或者异种（牛-羊）克隆胚胎在相同的培养系统下（B₂原装无血清培养液[法国CCD公司产品]+VERO细胞饲养层）进行体外培养。实验共进行7个批次。牛、羊的卵母细胞成熟率在相同体外成熟培养液内没有明显差异。但是克隆胚胎的体外培养情况差异极显著。培养同种（羊-羊）重组胚胎113枚,分裂数量为97枚,发育到桑囊胚数量为67枚;培养异种（牛-羊）克隆胚胎57枚,分裂数量为44枚,发育到桑囊胚数量为2枚。结果显示,在上述克隆胚胎培养系统下,同种（羊-羊）重组胚胎与异种（牛-羊）克隆胚胎发育能力存在极显著的差异。更多还原

【Abstract】 The goat and bovine oocytes are cultured in the IVM-medium containing TCM-199 with 0.22 mg/ml 10 mM Hepes, 0.1 IU/ml HMG and 1 μ g/ml E2,10% FCS(fetal calve serum), then the oocytes with visible PB1 are selected for enucleating and reconstructed cloning embryos with ear epithelia cells of Boer goat. The successfully fused embryos are cultured under free serum cultural systems (B2+VERO feeding cells) in vitro. After 48 h, 2-4 cells stage embryos in nomal morphology with the same size blastomeres are moved into new medium drop with B2 and VERO feeding cells for further development. The development rates of morula and blastocysts of the goat intraspecies cloned embryos are 59.3% (67/113), the development ratas of morurla and blastocysts of the bovine-goat interspecies embryos are 3.5%(2/57).The results suggest there are significant differences between the development ability of goat intraspecies and bovine-goat interspecies cloned embryo.更多还原