睾丸精子和附睾精子冷冻保存研究

Study on cryopreservation of testicular and epididymal spermatozoa

目的探讨以0.25ml麦管为载体,Irvine为精子冷冻保护剂的方法在睾丸精子和附睾精子冷冻复苏中的应用。方法收集2019年7月至2020年12月期间诊断性睾丸/附睾穿刺术后剩余的睾丸精子悬液样本10份和附睾精子悬液样本20份,以0.25ml麦管为载体,Irvine为精子冷冻保护剂进行冷冻复苏,评估冷冻复苏效果。结果冷冻前10份睾丸精子悬液样本活动精子数目为5~8条/(200HP,40个视野)。复苏后1份睾丸精子悬液样本活动精子数目为5条/(200HP,80个视野),5份睾丸精子悬液样本活动精子数目为3~4条/(200HP,80个视野),4份睾丸精子悬液样本活动精子数目为1~2条/(200HP,80个视野)。冷冻前20份附睾精子悬液样本精子活动率为(13.65±9.06)%,复苏后精子活动率为(6.85±4.61)%,显著性下降(t=6.521,P<0.001),复苏率为(49.88±7.96)%。结论以0.25ml麦管为载体,Irvine为精子冷冻保护剂的方法在睾丸精子和附睾精子冷冻复苏中有较好的复苏率,材料来源方便,冷冻复苏操作简便,储存空间小等优势适合实验室日常工作需要。

Objective To explore the application of 0.25ml straw as carrier and Irvine as sperm cryoprotectant in cryopresuscitation of testicular and epididymal spermatozoa.Methods 10 samples of testicular sperm suspension and 20 samples of epididymal sperm suspension after diagnostic testicular/epididymal puncture from July 2019 to December 2020 were collected.The cryoresuscitation was performed with 0.25ml straw as carrier and Irvine as sperm cryoprotectant to evaluate the effect of cryoresuscitation.Results The number of motile sperm in 10 testicular sperm suspension samples was 5~8/(200 HP,40 visual fields).After resuscitation,the number of motile sperm in one testicular sperm suspension sample was 5/(200HP,80 visual fields),the number of motile sperm in 5 testicular sperm suspension samples was 3~4/(200HP,80 visual fields),and the number of motile sperm in 4 testicular sperm suspension samples was 1~2/(200HP,80 visual fields).The sperm motility of 20 epididymal sperm suspension samples was(13.65±9.06)% before freezing and(6.85±4.61)% after resuscitation,which decreased significantly(t=6.521,P<0.001),and the recovery rate was(49.88±7.96)%.Conclusion The method of using 0.25ml straw as carrier and Irvine as sperm cryoprotectant has a good recovery rate in testicular and epididymal sperm cryoresuscitation.It has the advantages of convenient material source,simple cryoresuscitation operation and small storage space,which is suitable for the daily work of the laboratory.