Cryopreservation of rhesus monkey (Macaca mulatta) epididymal spermatozoa before and after refrigerated storage
Cryopreservation of rhesus monkey (Macaca mulatta) epididymal spermatozoa before and after refrigerated storage
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DOI：
10.2164/jandrol.107.003921
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发表时间：
2008-05-01
影响因子：
--
通讯作者：
Vandevoort, Catherine A.
中科院分区：
文献类型：
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作者：
Dong, Qiaoxiang;Rodenburg, Sarah E.;Vandevoort, Catherine A.
最近，人们对保存附睾精子作为遗传资源库的潜在材料来源越来越感兴趣;然而，尚未探索恒河猴附睾精子的冷冻保存。本研究评价了不同条件下长时间冷藏保存完整附睾尾部对猕猴附睾精子解冻后活力的影响，并测试了改变冷冻保护剂和冷却方法是否能提高冷冻保存后附睾精子的解冻后活力。冷冻前的运动能力在冷藏（0 ℃）24或48小时后显著下降。虽然冷冻储存24小时后解冻后活力没有显著差异，但在较高温度（4 ℃-10 ℃）下储存的附睾效果更好，但在4 ℃下冷冻储存48小时后解冻后活力仍显著下降。甘油和乙二醇在3%和6%的比较显示相似的解冻后运动。然而，在所有冷冻试验中，无论样品是新鲜采集还是冷藏储存24或48小时后采集，均使用3%甘油获得了一致的高解冻后活力。冷却在较高的速度为220摄氏度/分钟，发现产生更好的解冻后的运动比29摄氏度/分钟的较慢的速度。解冻时间进行了评估，并需要至少30秒的解冻0.25-mL吸管含有50 μ L精液样本。冷冻保存24或48小时后，用3%甘油包装并冷却的恒河猴附睾精子解冻后活力的总体平均值为42%。附睾精子解冻后活力的结果表明，这种方法应该是实用的保存附睾精子，即使组织必须从远离冷冻保存实验室的地点运送。
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 postthaw motility for epididymal sperm after refrigerated storage. Motility before freezing decreased significantly after refrigerated storage (O 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-mu L 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.