马鹿XY精子分选及冷冻精液制备试验

Cryopreservation of Flow-sorted X-or Y-chromosome Bearing Sperm on Wapiti Stags

为获得分选马鹿X、Y冷冻精液,对电刺激采集的15只马鹿精液经过7h长距离运输,流式细胞仪分离、冷冻,解冻后精液重分析和品质鉴定。结果显示,常规冻精和X、Y型冻精性别比分别为50%、93.11%和95.18%,常规冻精与X、Y型冻精组间差异显著(p<0.05);活率分别为0.45±0.04、0.55±0.04和0.53±0.03,常规冻精与X、Y型冻精组间差异不显著(p>0.05);存活指数分别为0.05、0.05和0.25,常规冻精与X、Y型冻精组间差异显著(p<0.05);顶体完整率分别为72.6±1.0%、74.6±0.5%和72.2±0.6%,常规冻精与X、Y型冻精组间差异不显著(p>0.05)。结论:马鹿XY分选冷冻精液性别比和品质达到性别控制人工授精的要求。

This study were to develop a method of production of frozen-thawed sorted X- or Y- chromosome beating sperm on wapiti deer. Methods： Semen were collected by electro-ejaculated from stags and were transported about 7h, and then were separated into X- and Y- chromosome bearing sperm after analysis and reanalysis with a modified high-speed cell sorter or unsorted and finally were frozen-thawed. Sorted sperm（x or y） and control sperm were Reanalysised and assessed. Results： the sex ratio of frozen-thawed sorted X- or Y- chromosome bearing sperm was superscripts differ （50%、93.11%、95.18%） than unsorted sperm of the control; the population of the sorted sperm with forward progressive motility was not significantly lower （0.55±0.04,0.53± 0.03）than that of unsorted sperm of the control （0.45± 0.04）; so did Post thaw acrosome integrity （74.6±0.5,72.2±0.6,72.6± 1.0）. Conclusion：this study has demonstrated it is possible to obtain sorted frozen-thawed wapiti stags sperm for AI using flow cytometry techniques.