Dong Qiaoxiang, Rodenburg Sarah E, Huang Changjiang, Vandevoort Catherine A
California National Primate Research Center, University of California, Davis, CA 95616, USA.
J Androl. 2008 May-Jun;29(3):283-92. doi: 10.2164/jandrol.107.003921. Epub 2008 Jan 9.
Recently, there has been an increased interest in preservation of epididymal sperm as a potential source of material for genetic resource banking; however, cryopreservation of epididymal sperm from the rhesus monkey has not been explored. This study evaluated the effect of prolonged refrigerated storage of the intact cauda epididymides at various conditions on the postthaw motility of rhesus monkey epididymal spermatozoa, and also tested whether altering cryoprotectants and cooling methods could improve post-thaw motility for epididymal sperm after refrigerated storage. Motility before freezing decreased significantly after refrigerated storage (0 degrees C) for a period of 24 or 48 hours. Although postthaw motility was not significantly different after 24 hours of refrigerated storage, epididymides stored at a higher temperature (4 degrees C-10 degrees C) yielded better results, but postthaw motility still decreased significantly after 48 hours of refrigerated storage at 4 degrees C. Comparisons of glycerol and ethylene glycol at 3% and 6% revealed similar postthaw motility. However, consistently high postthaw motility was obtained with 3% glycerol throughout all freezing trials regardless of whether samples were collected fresh or after refrigerated storage for 24 or 48 hours. Cooling at a higher rate of 220 degrees C/min was found to yield better postthaw motility than the slower rate of 29 degrees C/min. Thawing time duration was evaluated, and a minimum of 30 seconds was required for thawing 0.25-mL straws containing 50-microL semen samples. An overall average of 42% postthaw motility was obtained for rhesus monkey epididymal sperm packed in 3% glycerol and cooled after 24 or 48 hours refrigerated storage. These postthaw motility results for epididymal sperm indicate that this method should be practical for use in preserving epididymal sperm, even if tissue must be shipped from sites remote from the cryopreservation laboratory.
最近，人们对保存附睾精子作为遗传资源库潜在材料来源的兴趣日益增加；然而，恒河猴附睾精子的冷冻保存尚未得到探索。本研究评估了在不同条件下完整附睾尾部长期冷藏对恒河猴附睾精子解冻后活力的影响，还测试了改变冷冻保护剂和冷却方法是否能提高冷藏后附睾精子的解冻后活力。冷冻前的活力在冷藏（0℃）24或48小时后显著下降。虽然冷藏24小时后的解冻后活力没有显著差异，但在较高温度（4℃-10℃）下储存的附睾产生了更好的结果，但在4℃冷藏48小时后解冻后活力仍显著下降。3%和6%的甘油与乙二醇的比较显示解冻后活力相似。然而，在所有冷冻试验中，无论样本是新鲜采集还是冷藏24或48小时后采集，使用3%甘油始终能获得较高的解冻后活力。发现以220℃/分钟的较高速率冷却比29℃/分钟的较缓慢速率能产生更好的解冻后活力。对解冻时间进行了评估，解冻0.25mL含有50μL精液样本的细管至少需要30秒。对于包装在3%甘油中并在冷藏24或48小时后冷却的恒河猴附睾精子，解冻后活力的总体平均水平为42%。附睾精子的这些解冻后活力结果表明，即使组织必须从远离冷冻保存实验室的地点运送过来，这种方法在保存附睾精子方面应该也是可行的。
