冷冻保存对人类附睾精子功能的影响

Effect of cryopreservation on human epididymal sperm functions

目的：了解人类附睾精子经历冷冻保存后的精子功能变化。方法：应用精子功能检测方法。结果：１１ 例先天性双侧输精管缺如患者附睾精子标本冷冻保存后，每例冷冻标本均见活动精子，但精子活动率显著降低，精子活动能力减弱。精子膜完整率显著下降，头膜损伤－ 尾膜完整的精子率（ Ⅲ型精子） 显著升高，精子体外存活时间显著缩短。精子顶体蛋白酶活性降低。去透明带地鼠卵穿透试验检验精子体外授精能力失败的标本，采用显微注射技术可使精子辅助授精。结论：（１） 冷冻保存对附睾精子功能有显著的影响，造成精子功能减弱；（２） 冷冻精子仅用单一尾膜肿胀试验筛选，有可能拾取到的是尾膜完整，但头膜损伤的精子；（３） 建立显微授精技术可有效地对冷冻附睾精子辅助受精。

Aim:To investigate the effect of cryopresevation on human epididymal sperm functions. Methods: Epididymal sperm was obtained from men with congenital absence of the vas deferens ( n=11 ). Frozen sperm functions were examined by using motility evaluation, sperm head-tail membrane integrity test, survival time test, thin gelatin lager assay and zona-free hamster oocyte penetration test. Results:The results showed that sperm motility, motility grade, membrane integrity rate, survival time in vitro, acrosomal proteinase activity and fertilization rate had significantly lowered after thawing ( P<0 01 ). Using intracytoplasmic frozen epididymal sperm injection into hamster oocytes, swellen sperm head could be observed.Conclusion:Human sperm could undergo the freezing-thawing process. However, frozen sperm functions had significantly decreased. Damage to the head membrane and the tail could occur independently. Sperm injection would benefit the fertilization of frozen epididymal sperm.