Rhesus monkey sperm cryopreservation with TEST-yolk extender in the absence of permeable cryoprotectant.
Rhesus monkey sperm cryopreservation with TEST-yolk extender in the absence of permeable cryoprotectant.
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DOI：
10.1016/j.cryobiol.2008.09.014
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发表时间：
2009-02
期刊：
影响因子：
2.7
通讯作者：
VandeVoort, Catherine A.
中科院分区：
文献类型：
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作者：
Dong, Qiaoxiang;Correa, Liane M.;VandeVoort, Catherine A.
最近，人们对哺乳动物精子的超快速冷冻越来越感兴趣，特别是在没有冷冻保护剂的情况下进行玻璃化冷冻。非人灵长类动物的精子冷冻保存已经取得了成功，但在标准人工授精（AI）中使用冷冻-解冻精子仍然很困难，去除可渗透的冷冻保护剂可能会增加AI的成功率。本研究旨在探索在无渗透性冷冻保护剂的情况下冷冻恒河猴精子的可能性。具体而言，我们评估了各种因素，如蛋黄的存在或不存在，蛋黄在扩展剂中的百分比，以及冷却和解冻速率对无渗透性冷冻保护剂的冷冻成功的影响。结果显示，在没有蛋黄的情况下用TEST冷冻几乎没有提供保护（解冻后运动性<15%）。在试验中，40%或更多的蛋黄导致运动力降低，而在20-30%的范围内的蛋黄产生最活跃的精子。以缓慢速率（29 °C/min）冷却显著降低了仅用TEST-蛋黄冷冻的样品的解冻后运动性，但对甘油存在下的对照没有影响。类似地，在室温空气中缓慢解冻对于没有渗透性冷冻保护剂的冷冻是有害的（<2%运动性）。除了运动性，精子获能的能力基于cAMP和咖啡因激活后细胞内钙水平的增加，表明新鲜和冻融的运动精子之间没有差异，无论治疗如何。总之，本研究表明，当将样品加载到标准0.25 mL吸管中，在液氮蒸汽中以220 °C/min快速冷却，并在37 °C水浴中快速解冻时，在不存在渗透性冷冻保护剂的情况下，可以用TEST-蛋黄（20%）冷冻保存恒河猴的射精精子和附睾精子。这项研究也代表了非人类灵长类动物在没有渗透性冷冻保护剂的情况下首次成功冷冻。
Recently, there has been increased interest in ultra-rapid freezing with mammalian spermatozoa, especially for vitrification in the absence of cryoprotectants. Sperm cryopreservation in non-human primates has been successful, but the use of frozen–thawed sperm in standard artificial insemination (AI) remains difficult, and removal of permeable cryoprotectant may offer opportunities for increased AI success. The present study intended to explore the possibility of freezing rhesus monkey sperm in the absence of permeable cryoprotectants. Specifically, we evaluated various factors such as presence or absence of egg yolk, the percentage of egg yolk in the extenders, and the effect of cooling and thawing rate on the success of freezing without permeable cryoprotectants. Findings revealed that freezing with TEST in the absence of egg yolk offers little protection (<15% post-thaw motility). Egg yolk of 40% or more in TEST resulted in decreased motility, while egg yolk in the range of 20–30% yielded the most motile sperm. Cooling at a slow rate (29 °C/min) reduced post-thaw motility significantly for samples frozen with TEST-yolk alone, but had no effect for controls in the presence of glycerol. Similarly, slow thawing in room temperature air is detrimental for freezing without permeable cryoprotectant (<2% motility). In addition to motility, the ability of sperm to capacitate based on an increase in intracellular calcium levels upon activation with cAMP and caffeine suggested no difference between fresh and frozen–thawed motile sperm, regardless of treatment. In summary, the present study demonstrates that ejaculated and epididymal sperm from rhesus monkeys can be cryopreserved with TEST-yolk (20%) in the absence of permeable cryoprotectant when samples were loaded in a standard 0.25-mL straw, cooled rapidly in liquid nitrogen vapor at 220 °C/min, and thawed rapidly in a 37 °C water bath. This study also represents the first success of freezing without permeable cryoprotectant in non-human primates.
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影响因子：
--
作者：
Dong, Qiaoxiang;Rodenburg, Sarah E.;Vandevoort, Catherine A.
通讯作者：
Vandevoort, Catherine A.
影响因子：
0.7
作者：
Feradis, AH;Pawitri, D;Hayes, ES
通讯作者：
Hayes, ES
影响因子：
3.6
作者：
Isachenko, V;Isachenko, E;van der Ven, H
通讯作者：
van der Ven, H
影响因子：
2.8
作者：
Amirat, L;Tainturier, D;Anton, M
通讯作者：
Anton, M
DOI：
10.1080/01485010701225675
发表时间：
2007-01-01
期刊：
ARCHIVES OF ANDROLOGY
影响因子：
--
作者：
Hossain, A. M.;Osuamkpe, C. O.
通讯作者：
Osuamkpe, C. O.