不同冷冻方法对人睾丸精子冷冻保存效果的研究

摘要

Objective:To investigate the effectiveness of different cryoprotectant and carrier on testicular sperm cryopreservation.Methods:The patients had testicular biopsies to check whether there were any testicular sperms in the Reproductive Medicine Hospital of the First Hospital of Lanzhou University.After ICSI was performed on patients with mature motile sperm (n =46),the remaining testicular tissues were cryopreserved with an informed consent.Each testicular tissue was divided into four parts for freezing with different freezing methods:1.8 ml freezing tube + glycerol compound protective agent in group A,1.8 ml freezing tube + Fasrun cryoprotectant in group B,0.25 ml straw + Fasrun cryoprotectant in group C and ultra-trace freezing straw + Fasrun cryoprotectant in group D.The testicular tissues in each group was fumigated with liquid nitrogen,and then put into liquid nitrogen.The sperm viability and normal morphological rate after recovery were compared among the groups.Results:The motile sperms can satisfy the number of sperms for ICSI operations in each group.After thawing,the number of sample easy to find the normal motile sperm(＞0.1/200× field of view)in group C and D was significantly higher than that in group A and group B(P＜0.05).The recovery rate of testicular sperm in group D was significantly higher than that in group A and group B(P＜0.05),and the recovery rate in group C was significantly higher than that in group A(P＜0.05).Conclusions:The improved ultra-trace freezing straw plus Fasrun cryoprotectant can improve the resuscitation rate of testicular sperm.%目的 比较不同冷冻保护剂和不同冷冻载体对人睾丸精子的冷冻保存效果. 方法 选取在兰州大学第一医院生殖医学专科医院进行诊断性睾丸穿刺获得成熟活动精子的患者,以及行睾丸精子ICSI术后的患者,在知情同意下将剩余睾丸组织冷冻保存,共46例.将每例睾丸组织分为4份,随机分成4组进行冷冻:1.8 ml冷冻管+甘油复合冷冻剂(A组,46例);1.8 ml冷冻管+Fasrun冷冻保护剂(B组,46例);0.25 ml麦管+Fasrun冷冻保护剂(C组,46例);超微量冷冻麦管+ Fasrun冷冻保护剂(D组,46例).冷冻标本经液氮熏蒸后投入液氮,比较各组复苏后精子活力和复苏率. 结果 冷冻前各组的活动睾丸精子能够满足ICSI操作所用精子量;冷冻复苏后,C组和D组能够容易找到正常活动精子(≥0.1/200倍视野)的例数显著高于A组、B组(P＜0.05);D组解冻后运动精子复苏率显著高于A组、B组(P＜0.05),C组解冻后运动精子复苏率显著高于A组(P＜0.05). 结论 应用改良的超微量冷冻麦管+ Fasrun冷冻保护剂能够提高睾丸精子的冷冻复苏率.