人类精液一步熏蒸法冻融的实验研究

目的:研究一步熏蒸法对冷冻精子复苏率的影响,以探索人类精子的最优冷冻方法。方法:选择100例符合WHO第5版正常参考值的精液标本,按照程序降温法和一步熏蒸法分2组进行冷冻。利用计算机辅助精液分析系统(CASA)检测冷冻前后精子前向活动力,同时比较2种方法对精子膜功能完整性、畸形率的影响。结果:冷冻后程序降温法和一步熏蒸法前向运动精子百分率差异有显著性(34.0±18.4)%vs(43.0±19.5)%,P<0.05],均较冷冻前的(57.0±16.7)%显著下降(P<0.05);2种冷冻方法前向运动精子复苏率差异显著(52.2±20.6)%vs(67.1±20.0)%,P<0.05];与冷冻前(84.5±7.5)%]相比,2种方法冷冻后低渗肿胀精子百分率明显下降分别为(67.1±11.1)%和(70.6±10.0)%,P<0.05],2种冷冻方法间差异也显著(P<0.05);而畸形率冷冻后显著增加(85.0±8.7)%和(85.7±9.1)%vs(77.8±9.6)%,P<0.05],但2种冷冻方法之间差异无显著性(P>0.05)。结论:与程序降温法相比,一步熏蒸法操作简单方便,适用于不同时间段的精液标本冷冻,具有较高的冷冻复苏率,是一种值得推荐的快速冷冻方法。

Direct fumigation for freeze-thawing of human sperm: an experimental study

Objective:To study the effect of direct fumigation on the post-thaw recovery rate of cryopreserved spermatozoa,and to search for a best method for human sperm cryopreservation.Methods:We collected semen samples from 100 donors conforming to the normal reference values in WHO Laboratory Manual for the Examination and Processing of Human Semen(5th Ed),divided them into two groups,and subjected them to cyopreservation by programmable freezing(Group A) and direct fumigation(Group B),respectively.We detected the progressive motility of pre-freezing and post-thaw sperm with a computer-assisted semen analyzer,and compared the effects of the two methods on the functional integrity of sperm membrane and the rate of abnormal sperm using the precentage of hypo-osmotic sweelling sperm and modified Papanicolaou staining.Results:Statistically significant differences were found in post-thaw sperm progressive motility between the Groups A and B(34.0±18.4]% vs 43.0±19.5]%,P<0.05),both remarkably decreased as compared with pre-freezing(57.0±16.7]%,P<0.05).Such differences were also found in the post-thaw recovery rate of progressively motile sperm between the two groups(52.2±20.6]% vs 67.1±20.0]%,P<0.05).The post-thaw precentage of hypo-osmotic sweelling sperm was obviously decreased in both Groups A and B(67.1±11.1]% and 70.6±10.0]%) in comparison with pre-freezing(84.5±7.5]%,P<0.05),with significant differences between A and B(P<0.05).However,the rate of sperm abnormality was evidently increased in Groups A and B(85.0±8.7]% and 85.7±9.1]%),significantly higher than pre-freezing(77.8±9.6]%,P<0.05),but with no significant differences between A and B(P>0.05).Conclusion:Direct fumigation is superior to programmable freezing for its easier operation,wider application,and higher sperm recovery rate.