Tulane National Primate Research Center, Covington, LA 70433, USA.
Theriogenology. 2009 Oct 1;72(6):834-40. doi: 10.1016/j.theriogenology.2009.06.002. Epub 2009 Jul 30.
To determine whether sperm derived from the vas deferens could be retrieved and successfully cryopreserved, testes were collected from 20 rhesus macaques (Macaca mulatta). The males ranged in age from 3 to 19 yr with an average age of 8.5 yr. No sperm was obtained from three animals that were younger than 4 yr. The remaining 17 samples contained sperm with an average sperm cell number of 421.8+/-88.7x10(6) and an average motility of 72.8+/-4.4%. After 24h of culture in TALP medium at 37 degrees C in 5% CO(2) and 95% air, the overall motility decreased significantly in all samples regardless of treatment. Freezing in TEST (TES-Tris buffer)-yolk buffer containing 6% (vol/vol) glycerol had a significant effect on sperm, reducing the immediate postthaw motility to 42.4% in nontreated samples. Treatment with dibutyryl-cAMP and caffeine further reduced sperm motility after 4h in fresh sperm (72.8% vs. 50.4%) but increased motility in sperm that had been frozen (14.0% vs. 23.2%). The age of the male did not influence sperm concentration or grade but proved to be a significant factor in determining motility of frozen-thawed treated sperm, with lower motility found in samples from older males. Overall, the study demonstrates that motile sperm can be obtained from postmortem males, although subsequent studies will be needed to determine whether the quality is sufficient to facilitate its use in assisted reproduction.
为了确定是否可以从输精管中获取并成功冷冻保存精子,我们从 20 只恒河猴(Macaca mulatta)中收集了睾丸。雄性年龄从 3 到 19 岁不等,平均年龄为 8.5 岁。年龄小于 4 岁的 3 只动物没有获得精子。其余 17 个样本中含有精子,平均精子数量为 421.8+/-88.7x10(6),平均活力为 72.8+/-4.4%。在 37°C、5%CO(2)和 95%空气的 TALP 培养基中培养 24 小时后,所有样本的总活力无论是否处理都显著下降。在含有 6%(体积/体积)甘油的 TEST(TES-Tris 缓冲液)-卵黄缓冲液中冷冻对精子有显著影响,未经处理的样本中立即解冻后的活力降低至 42.4%。用二丁酰环腺苷酸和咖啡因处理后,新鲜精子的活力在 4 小时后进一步降低(72.8%对 50.4%),但冷冻精子的活力增加(14.0%对 23.2%)。雄性的年龄不会影响精子浓度或等级,但被证明是决定冷冻解冻处理精子活力的重要因素,年龄较大的雄性样本的活力较低。总的来说,这项研究表明,虽然还需要进一步的研究来确定其质量是否足以促进其在辅助生殖中的应用,但可以从死后的雄性中获得有活力的精子。