ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (9): 1315-1319.doi:

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Study on Handmade Somatic Cell Cloning in Dairy Cattle

ZHANG Nuo 1 , DU Weihua 1, HAO Haisheng 1,WANG Dong 1, WU Shen 1,2 ,ZHU Huabin 1*

  

  1. 1. Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193,China;2.Grassland Research Institute,Chinese Academy of Agricultural Sciences,Hohhot 010010,China
  • Received:2008-04-03 Revised:1900-01-01 Online:2009-09-24 Published:2009-09-24
  • Contact: ZHU Huabin

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Abstract:

To optimize the technique of handmade somatic cell cloning in dairy cattle, based on bovine oocytes, the different alternating current (AC) voltage, different activation methods of reconstructed embryos and different collection methods of cumulusoocyte complexes(COCs) were used. The results showed that: (1)The fusion rate of oocytes increased significantly with the decreasing of AC voltage（P<0.05）. (2)The cleavage rates and blastocyst rates of reconstructed embryos treated with A23187 + 6DMAP was significantly higher than that treated with ionomycin + 6DMAP（P<0.05）. (3)The total number of oocytes collected by negativepressure aspiration and by cutting ovaries was significantly higher than that collected by injector aspiration（P<0.05）, and the proportion of COCs of grade 12 got by cutting ovaries was higher（P<0.05）. In conclusion，using lower AC voltage，choosing A23187 + 6DMAP for reconstructed embryos activating,aspirating COCs with negativepressure aspiration can significantly improve the handmade somatic cell cloning efficiency of dairy cattle.