畜牧兽医学报 ›› 2006, Vol. 37 ›› Issue (12): 1287-1292.doi:

• 遗传繁育 • 上一篇    下一篇

鲁西黄牛体细胞克隆保种技术的研究

朱化彬;程金华;孙秀柱;王栋;戴蕴平;龚国春;万荣;郝海生   

  1. 1.中国农业科学院北京畜牧兽医研究所,北京 100094;2.中国农业大学农业生物技术国家重点实验室, 北京100094
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-12-25 发布日期:2006-12-25

Study on Somatic Cell Clone and Conservation Technique of Luxi Cattle

  • Received:1900-01-01 Revised:1900-01-01 Online:2006-12-25 Published:2006-12-25

摘要: 以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体,研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛(1♂,4♀)成纤维细胞系,作为核移植供体细胞,利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植,试验结果表明:重构胚的融合率为62.5%(242/387),分裂率为63.6%(154/242),体外培养第7天囊胚发育率为42.9%(66/154)。体外培养第7天的囊胚的内细胞团细胞(ICM)与滋养层细胞数平均为37和47,ICM占44.2%。体外发育到7 d的囊胚新鲜胚胎的移植妊娠率(新鲜胚胎)移植受体10头,60 d妊娠率为20%(2/10)。

Abstract: In order to conserve our native excellent yellow cattle breeds, the somatic cell clone technique was studied, in which the bovine ear skin fibroblast in vitro cultured was used as nuclear donor. The fibroblast line of Luxi cattle (4 females and 1 male) for nuclear donor was established through tissue pieces culture method, and the oocytes of ovaries collected from slaughter house were cultured for nuclear transfer receptor. Experiment results indicate that the fusion rate and cleavage rate of reconstruction embryos were 62.5% (242/387) and 63.6%(154/242) respectively, and the development rate of blastocysts was 42.9%(66/154) after culture 7 days. The average cell number of inner cell mass and trophoblast was 37 and 47 respectively, in which 44.2% was inner cell mass. 10 blastocyts of reconstruction embryo were transferred to 10 recipient after cultured 7days in vitro, the pregnancy rate was 20%(2/10).