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KMID : 0359319970370030639
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Korean Journal of Veterinary Research
1997 Volume.37 No. 3 p.639 ~ p.645
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Studies on the cloning of calves by nuclear transplantation II. Efficient embryo cloning under oocyte activation, cell cycle regulation of donor nuclei and optimal culture conditions
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Hwang Woo-Suk
Roh Sang-Ho
Lee Byeong-Chun
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Abstract
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The objectives of the present study were improvements in the efficiency of developmental rates to morula and blastocyst stages to produce a large number of genetically identical nuclear transplanted embryos. The oocytes collected from slaughterhouse ovaries were matured 24h in TCM199+10% FBS and exposed to $39℃ or room temperature to allow cytoplasmic maturation and gain activation competence. Donor embryos were treated for 12h with 10㎍/ml nocodazole or 0.05㎍/ml demicolcine to synchronize the cell cycle stage at 26h after the onset of culture. The blastomeres and recipient oocytes were fused by electrofusion. The cloned embryos were then cultured in various conditions to allow further development. In the treatment of oocyte activation and cell cycle regulation of donor nuclei, the room temperature exposure and nocodazole treatment group had significant effect on the developmental rates to morula/blastocyst(21.7% vs 12.1~16.7%), but had no significant effect on the fusion rates between donor blastomeres and recipient oocytes. The developmental rates of bovine nuclear transplanted embryos appeared to be higher significantly in mTALP medium under 5% O2 condition and in TCM199 with bovine oviduct epithelial cell under 20% O2 condition(22.2%) than other groups. In embryo transfer of nuclear transplanted embryos, there were no significant differences in calving rates between the use of excellent and good grade donor embryos.
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KEYWORD
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bovine , nuclear transplantation , activation , cell cycle
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