Grupo de Ecología y Biología de la Reproducción, Museo Nacional de Ciencias Naturales (CSIC), Madrid, Spain.
Theriogenology. 2009 Dec;72(9):1268-77. doi: 10.1016/j.theriogenology.2009.07.023. Epub 2009 Sep 22.
Postmortem sperm recovery from the epididymides may constitute a powerful tool for the conservation of valuable genetic material. The domestic cat (Felis catus) is a good model for wild felids and, using this model, we have explored the effect of epididymides storage time on sperm motility and percentage of intact acrosomes upon sperm recovery and after cryopreservation. We also examined the effect of time of sperm equilibration with glycerol before freezing on sperm motility and the percentage of intact acrosomes. Motility varied between sperm recovered from epididymides that were stored for different times. Significant differences were seen in the sperm motility index (SMI) before freezing (55.91+/-2.02, 48.21+/-1.47, and 43.03+/-1.32) and after thawing (51.81+/-3.02, 41.90+/-2.14, and 42.35+/-1.95) of sperm recovered from epididymides stored for 0, 48, or 72 h, respectively. The percentage of intact acrosomes did not vary significantly with storage time (average 60.33+/-1.38% before and 52.50+/-1.91% after freezing, respectively). The percentage of normal sperm after different storage times did not differ (average 19.22+/-1.25% normal sperm after recovery). When epididymides were stored for 72 h, time of sperm equilibration with glycerol (30 vs. 120 min) resulted in significant differences in both motility (SMI=39.17+/-2.76 and 45.00+/-2.65, respectively) and the percentage of intact acrosomes (45.76+/-4.91% and 60.67+/-3.64%, respectively) after thawing. In conclusion, best results are achieved when sperm are recovered from epididymides within 24h of cool storage and when they are equilibrated with glycerol during 120 min before freezing. The current results should be useful in the further development of techniques for the rescue and cryostorage of epididymal spermatozoa of endangered felids.
从附睾中回收死后精子可能成为保存有价值遗传物质的有力工具。家猫（Felis catus）是野生猫科动物的良好模型，我们利用该模型探讨了附睾储存时间对精子活力和冷冻前精子回收率及完整顶体率的影响。我们还研究了在冷冻前用甘油平衡精子时间对精子活力和完整顶体率的影响。从储存不同时间的附睾中回收的精子的活力存在差异。冷冻前的精子运动指数（SMI）存在显著差异（55.91+/-2.02、48.21+/-1.47 和 43.03+/-1.32），解冻后的精子运动指数（SMI）也存在显著差异（51.81+/-3.02、41.90+/-2.14 和 42.35+/-1.95），分别为从储存 0、48 或 72 小时的附睾中回收的精子。冷冻前后完整顶体率差异不显著（冷冻前平均为 60.33+/-1.38%，冷冻后平均为 52.50+/-1.91%）。不同储存时间的正常精子比例无差异（平均为 19.22+/-1.25%）。当附睾储存 72 小时时，精子与甘油平衡时间（30 分钟与 120 分钟）对解冻后的活力（SMI=39.17+/-2.76 和 45.00+/-2.65）和完整顶体率（45.76+/-4.91%和 60.67+/-3.64%）均有显著影响。总之，最好的结果是在附睾冷却储存 24 小时内从附睾中回收精子，并在冷冻前用甘油平衡 120 分钟。这些结果对濒危猫科动物附睾精子的拯救和冷冻保存技术的进一步发展具有重要意义。
