Structural integrity and developmental potential of spermatozoa following microwave-assisted drying in the domestic cat model
Structural integrity and developmental potential of spermatozoa following microwave-assisted drying in the domestic cat model
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DOI：
10.1016/j.theriogenology.2017.07.037
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发表时间：
2017-11-01
期刊：
影响因子：
2.8
通讯作者：
Comizzoli, Pierre
中科院分区：
文献类型：
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作者：
Patrick, Jennifer L.;Elliott, Gloria D.;Comizzoli, Pierre
表征哺乳动物细胞对非生理条件的恢复能力对于制定低温或环境温度下的保存和长期储存策略是必要的。使用家猫模型，该研究的目的是表征微波辅助干燥至与超零温度下玻璃态储存相容的水分含量后精子的结构完整性（形态和 DNA 损伤）以及功能特性（精子注射后的精子紫苑形成和胚胎形成）。在实验 1 中，用溶血素对猫附睾精子进行穿孔，并在海藻糖存在下在低湿度环境 (11%) 下干燥（使用设置为 20% 功率的商用微波炉）长达 50 分钟，然后测量水分含量和样品温度。在实验 2 中，对干燥长达 30 分钟的精子（使用与上述相同的方法）与新鲜精子的形态和 DNA 完整性进行了评估。在实验 3 中，根据精子紫苑形成（注射后 5 小时）和体外 7 天的胚胎发育，对注射到卵母细胞后干燥 30 分钟的精子与新鲜精子细胞的功能进行了评估。微波辅助干燥 30 分钟后，达到与干燥状态储存相容的水分含量。再水化后，精子形态未受影响，DNA 受损细胞的百分比（约 6.5%）与新鲜对照相似。与新鲜对照相比，干燥再水化的细胞的精子直径似乎通常更小。这一观察结果与注射干燥精子（6.5%）后囊胚形成比例低于新鲜精子（15%）相一致。然而，基于总卵裂球数量的囊胚质量不受精子处理的影响。这是迄今为止在任何物种中第一个令人鼓舞的报告，证明可以使用微波干燥精子，而不会对细胞结构和功能造成不可逆转的损害。由爱思唯尔公司出版
Characterizing the resilience of mammalian cells to non-physiological conditions is necessary to develop preservation and long-term storage strategies at low or ambient temperatures. Using the domestic cat model, the objective of the study was to characterize structural integrity (morphology and DNA damage) as well as functional properties (sperm aster formation and embryo formation after sperm injection) of spermatozoa after microwave-assisted drying to a moisture content compatible with storage in a glassy state at supra-zero temperatures. In Experiment 1, cat epididymal spermatozoa were porated with hemolysin and dried (using a commercial microwave oven set to 20% power) in the presence of trehalose for up to 50 min in a low humidity environment (11%) before measuring moisture content and sample temperature. In Experiment 2, morphology and DNA integrity were evaluated in sperm dried for up to 30 min (using the same method as above) versus fresh spermatozoa. In Experiment 3, the functionality of sperm dried for 30 min versus fresh sperm cells was evaluated after injection into oocytes based on sperm aster formation (5 h post-injection) and embryo development in vitro over 7 days. Moisture contents compatible with dry state storage were reached after 30 min of microwave-assisted drying. After rehydration, sperm morphology was not affected and the percentages of cells with damaged DNA (similar to 6.5%) was similar to the fresh controls. Sperm aster diameters appeared to be generally smaller for dried-rehydrated cells compared to the fresh controls. This observation was consistent with a lower proportion of blastocyst formation after injection with dried spermatozoa (6.5%) compared to fresh spermatozoa (15%). However, the blastocyst quality based on the total blastomere number was not affected by the sperm treatment. This is the first and encouraging report in any species so far demonstrating that spermatozoa can be dried using microwaves without causing irreversible damage to the cellular structure and function. Published by Elsevier Inc.