不同培养液对蓝狐精子体外获能效果的比较试验

采用上游法分离优化精子,用mTyrod’s液(T)、BO液(B)以及高渗液(HIS)3种不同培养液,在5%CO2的培养箱中进行获能培养,获能培养时间为6h。利用考马斯亮蓝染色法检测精子顶体反应率,伊红-苯胺黑染色法检测活精子比率以及观测法检测精子活力。在培养的0、1、2、4、6h时分别检测上述指标并统计分析,从而筛选出适合蓝狐精子体外获能的培养液。结果显示:B培养液优于T、HIS培养液,最佳获能培养时间t≥6h;HIS可以提高精子顶体反应率,但较高的渗透压不利于精子保存,不建议使用。

Different Culture Fluid on Capacitation in Vitro of Alopex lagopus Sperm

Sperm capacitation in vitro is the basic process of in vitro fertilization（IV）,and it is very significant to the reproduction study and embryo engineering.The swim-up method was used for selecting sperms in this experiment.The fresh ejaculated semen of blue fox was incubated with 3 kinds of substrates：mTyrod＇s solution（T）,BO solution（B）and the high ionic strength medium（HIS）at 38.5 ℃in 5% CO2 incubation box.The incubation time was 6 hours.The acrosome reaction rate was determined by coomassie blue staining and sperm viability was determined by eosin-nigrosine staining.Sperms activity was observed at 0,1,2,4 and 6 hour.Based on the statistical analysis of data,the most suitable culture system for sperm capacitation of blue fox can be selected.The results showed that BO solution（B）was better than mTyrod＇s solution（T）and high ionic strength medium（HIS）;the incubation time suitable for capacitation was t≥6 hour;the high ionic strength medium can increase acrosome reaction rate,but it was not good for sperm preservation.