不同甘油浓度与平衡时间对食蟹猴精液冷冻效果的影响

Effects of Glycerol Concentration and Equilibration Time on Cryopreservation of Cynomolgus Monkey Spermatozoa

探讨了不同甘油浓度（3%、5%、7%、11%）和不同平衡时间（30、60、90、120min）对食蟹猴（Macaca fascicularis）精液冷冻效果的影响,以建立和优化食蟹猴精液冷冻的程序。参照TTE稀释液成分组成改良型TTE,冷冻前和解冻后均检测精子的活力、畸形率、质膜完整性、顶体完整率。结果显示,平衡时间为30min时精子的冷冻解冻后活力、复苏率均高于平衡时间90min和120min组,差异显著（P〈0.05）,比60min组稍好;甘油浓度为3%、5%组的精子冷冻解冻后活力及复苏率均高于甘油浓度11%组,差异显著（P〈0.05）,比7%组好;不同甘油浓度各组间以及不同平衡时间各组间畸形率、质膜完整性、顶体完整率差异不显著（P〈0.05）。由此得出如下结论,在食蟹猴精液冷冻中,在改良TTE中加入3%-5%的甘油且平衡30min可以获得较好效果,精子冻后活率和复苏率达到45%和62%。

In order to optimize the protocols for Cynomolgus Monkey（Macaca fascicularis）sperm cryopreservation,the effects of glycerol concentration [3%,5%,7% and 11%（v/v）] and equilibration time（10,30,60 and 90 min）on cryopreservation of spermatozoa of Cynomolgus Monkey were studied.A modified TTE（mTTE）dilution solution was used and the sperm motility,morphology,membrane integrity and acrosome integrity were examined before freezing and post-thawing.When 30 min equilibration was applied,the post-thawing motility and recovery rates was significantly higher than those when 90 min or 120 min equilibration（P〈0.05）.The effect of 30 min equilibration was also better than that of 60 min equilibration.The post-thawing motility and recovery rate of spermatozoa was significantly higher（P〈0.05）for 3% and 5% glycerol groups than those for 11% glycerol group,and the effect of 3% and 5% glycerol was also better than 7% glycerol.The different glycerol concentrations and equilibration time showed similar effects on the membrane integrity,acrosomal integrity and malformation rate（P〈0.05）.The results suggest that addition of 3% or 5% glycerol and 30 min equilibration are beneficial for freezing Cynomolgus Monkey spermatozoa.