| In the research, two different methods (aspiration and incision) were evaluated for efficiency of equine oocytes collection. Two maturation media were used for in vitro maturation of equine oocytes. In medium 1, the basic medium was M199; in Medium 2, the basic medium was DMEM/F12.The results shown that incision method would be batter than aspiration method for efficiency of equine oocytes collection (83%vs46.5%)(p<0.05). There have no significant differences for the maturation rate observed among two different maturation media. The maturation rate of equine oocytes matured in medium M199 was 41.7% for Cp oocytes and 64.7% for Ex oocytes; in medium DMEM/F12 those were 46.7% and 66.7%(p>0.05). The maturation rate of Ex oocytes were higher in medium M199 than in medium DMEM/F12,but have no significant differences(p>0.05). The MII stage Cp oocytes cultured in different culture medium were activated by respectively exposure to inomycin 5 min followed by an incubation for 4 h in 2 mM/ml 6-DMAP and 10 ug/ml CHX, the percentage of cleaved have no significant differences, it was 45% in M199 medium and 57.1% in DMEM/F12 medium(p>0.05).Four different passages (F3, F6, F9, F15) of the fibroblasts were used as nuclear donors for the experiment of somatic cell nuclear transfer, the result showed that the cleavage rate of reconstructed embyro was not significantly different (P> 0.05).In the fusion emperiment,three fusion voltage respectively 1.8KV/cm, 2.0KV/cm and 2.4KV/cm, pulse duration 20μs, pulse frequency 2, were eused,when the fused voltage were 2.0 kV / cm and 2.4kV/cm, the fusion rates were 53.3% and 50%, their have no significant difference, but were significantly higher than the fusion rate of 29.2% which was gained when the fuson voltage was 1.8kV/cm. |
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