中国农业科学 ›› 2004, Vol. 37 ›› Issue (10): 1550-1554 .

• 畜牧.兽医.资源昆虫 • 上一篇    下一篇

体细胞克隆牛超排胚胎玻璃化冷冻及移植试验

董雅娟,柏学进,李建东,程明   

  1. 山东省莱阳农学院动物胚胎工程中心
  • 收稿日期:2004-02-04 修回日期:2004-06-24 出版日期:2004-10-20 发布日期:2004-10-20
  • 通讯作者: 董雅娟

The Experiment on the Cloned Cattle Derived from Somatic Cells and the Transfer of the Vitrified-Thawed Embryos of the Cloning Cattle

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  1. 山东省莱阳农学院动物胚胎工程中心
  • Received:2004-02-04 Revised:2004-06-24 Online:2004-10-20 Published:2004-10-20

摘要: 对体细胞克隆牛进行了超数排卵、胚胎玻璃化冷冻及胚胎移植试验。首先对体外受精卵进行了玻璃化冷冻保存试验,结果表明,体外受精囊胚经过EPS10和EPS20 的TCM-199液玻璃化冷冻-解冻后胚胎形态正常率和发育率各组间差异不显著,囊胚孵出率EPS20组为31.3% (35/112)极显著高于EPS10组的12.2% (13/107)(P<0.01),选择玻璃化冷冻液EPS20作为体细胞克隆牛超排采卵获得胚胎的冷冻保存液。本试验对体细胞克隆牛"康康"和"双双"进行了超排试验,分别获得6枚(4枚可用胚)和10枚(9枚可用胚)胚胎,各取2枚1级胚胎应用EPS20玻璃化液进行玻璃化冷冻,解冻后分别移植到4头同期发情处理的中国荷斯坦奶牛的黄体侧子宫角内,结果为1头9908号受体母牛妊娠,并且产下1头健康的体细胞克隆牛"双双"的胚胎移植后代。试验结果表明,①体细胞克隆牛与正常牛一样对外源激素(FSH)具有较好的应答能力和超数排卵的能力;②体细胞克隆牛的胚胎可用于玻璃化冷冻保存。

关键词: 克隆牛, 超数排卵, 胚胎, 玻璃化冷冻

Abstract: In this experiment, it was designed to carry out superovalation on the two cloned cattle, vitrification and transfer of the embryos recovered from them. First of all, it was carried out vitrification on embryos obtained by IVF. Results showed that there was no significant difference between the blastocysts (obtained by IVF) vitrified in EPS10 and these in EPS20 in the resuscitative rate and the developmental rate. The hatched rate of the blastocysts vitrified in EPS10 (31.3%,35/112) was significantly higher than that in EPS20(12.2%,13/107)(P<0.01), so EPS20 was selected as the vitrification solution to freeze the embryos recovered from the cloned cattles. After superovulation, six (four usable embryos) and ten (nine usable embryos ) embryos were respectively recovered from "Kang kang"and "Shuang shuang". Two embryos were selected from the recovered embryos of each cloned cattle to freeze in EPS20, subsequently thawed and transferred into ipsilateral horns of 4 Holstein recipient cows after synchronization of estrus, respectively. At last, one recipient cow (No 9908) became pregnant and delivered one healthy calf (descendant of the cloned cattle-"Shuang Shuang").The results of this experiment shows that the cloned cattle as well as common cattle had better response to the exotic FSH and better ability to multiovulation, the embryos recovered from the cloned cattle can be vitrificated.

Key words: Cloned cattle, Superovulation, Embryo, Vitrification