This study was carried out to investigate the optimal activation condition for parthenogenetic development. In order to activate oocytes at 24 hrs post onset of maturation, the oocytes were cultured 3∼13 μM Ca^2+ for 5 min, 5-8 ㎍/㎖ cytoclacin for 6 hrs, 0.5∼2.0 mM 6-dimethylaminopurine(DMAP) for 3 hrs alone or combination. The activated oocytes were clutured in TCM-199 media at 5% CO_2, 95% air, 38℃.
The cleavage rate after 48 hrs culture of oocytes treated with 3-13 μM Ca^2+, 5-8 ㎍/㎖ cytoclacin and 0.5∼2.0 mM DMAP for 5 min, 6 hrs and 3 hrs were 9.6%∼20.0%, 0.0%∼7.3% and 9.4%∼21.8%, 0.0%∼7.3% and 9.1%∼21.8% and 0.0%∼7.3%, respectively. When oocyte were treated with 10 μM Ca^2+, 10 ㎍/㎖ cytoclacin and 2.0 mM DMAP the blastocyst formation rate was significantly higher than other group. The cleavage rate after 48 hrs cultrue of oocytes treated with Ca^2+＋cytoclacin, Ca^2+ + DMAP, cytoclacin + DMAP were 75.9%∼93.5% and 9.7%∼19.0%, respectively. When oocytes were treated with Ca^2+ followed by DMAP, the blastocyst formation rate was significantly higher than other group(p＜0.05). When necleus transferred embryos co-cultured with bovine serum albumin(BSA), epithemal growth factor(EGF) and calf serum(CS), the developmental rate to blastocyst were higher than control group.
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