Bernklau Elisabeth, Wehrend Axel, Farshad Abbas
Veterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University of Giessen, 35392 Giessen, Germany.
Vet Sci. 2025 Aug 31;12(9):840. doi: 10.3390/vetsci12090840.
(1) Background: Cryopreservation of epididymal spermatozoa in dogs is challenging due to their lower cryotolerance compared to ejaculated spermatozoa. Given the limited sperm volume obtained per individual, efficient recovery and preservation techniques are essential. (2) Methods: This study assessed sperm collection and cryopreservation methods from the cauda epididymis of ten dogs undergoing routine elective castration. After dissection and mincing, the cauda epididymidis tissue was incubated in 0.9% saline at 38 °C for either 10- or 30-min. Samples were analyzed for concentration and motility using AndroVision software (CASA; AndroVision™; Minitüb GmbH) (Tiefenbach, Germany). Additional evaluations included histological examination, hypoosmotic swelling test, live/dead staining, and morphological assessments. Three extenders, custom-made Tris-Fructose-Citrate (Tris), custom-made Uppsala, and commercial Optixcell, were used for cryopreservation and compared for post-thaw sperm quality. (3) Results: No significant differences were found between the 10- and 30-min incubation groups regarding sperm motility, viability, or histological integrity. The total sperm counts were 292 × 10 ± 175 × 10 for the 10 min group and 233 × 10 ± 162 × 10 for the 30 min group ( = 0.56). Histological sections revealed no significant difference in residual intraluminal spermatozoa between groups, indicating that 10 min of incubation is sufficient for effective sperm migration. Post-thaw sperm motility was significantly higher with Uppsala (17.2 ± 12.2%) and Optixcell (11.7 ± 6.5%) compared to Tris (4.7 ± 4.8%). Morphological abnormalities were lowest in samples preserved with Optixcell (37.5 ± 10.1%, = 0.005). (4) Conclusion: A 10 min incubation period is adequate for efficient recovery of epididymal sperm in dogs. Among the tested extenders, Uppsala and Optixcell demonstrated superior cryoprotective effects, resulting in better post-thaw motility and reduced morphological abnormalities compared to Tris.
(1) 背景：与射出的精子相比，犬附睾精子的冷冻耐受性较低，因此犬附睾精子的冷冻保存具有挑战性。鉴于从每只犬获得的精子体积有限，高效的回收和保存技术至关重要。(2) 方法：本研究评估了对十只接受常规择期去势手术的犬的附睾尾进行精子采集和冷冻保存的方法。解剖并切碎后，将附睾尾组织在38℃的0.9%盐水中孵育10分钟或30分钟。使用AndroVision软件（计算机辅助精子分析系统；AndroVision™；Minitüb GmbH）（德国蒂芬巴赫）分析样本的浓度和活力。其他评估包括组织学检查、低渗肿胀试验、活/死染色和形态学评估。使用三种稀释液，即定制的Tris-果糖-柠檬酸盐（Tris）、定制的乌普萨拉液和商业用Optixcell液进行冷冻保存，并比较解冻后精子质量。(3) 结果：在精子活力、存活率或组织学完整性方面，10分钟和30分钟孵育组之间未发现显著差异。10分钟组的总精子数为292×10±175×10，30分钟组为233×10±162×10（P = 0.56）。组织学切片显示两组之间管腔内残留精子无显著差异，表明10分钟的孵育足以实现有效的精子迁移。解冻后，与Tris液（4.7±4.8%）相比，使用乌普萨拉液（17.2±12.2%）和Optixcell液（11.7±6.5%）时精子活力显著更高。使用Optixcell液保存的样本中形态异常最低（37.5±10.1%，P = 0.005）。(4) 结论：10分钟的孵育期足以有效回收犬的附睾精子。在所测试的稀释液中，乌普萨拉液和Optixcell液表现出更好的冷冻保护效果，与Tris液相比，解冻后活力更高，形态异常减少。
