Single sperm cryopreservation on cryoloops: an alternative to hamster zona for freezing individual spermatozoa
Single sperm cryopreservation on cryoloops: an alternative to hamster zona for freezing individual spermatozoa
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DOI：
10.1016/s1472-6483(10)62109-8
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发表时间：
2004-07-01
影响因子：
4
通讯作者：
Goldfarb, J
中科院分区：
文献类型：
--
作者：
Desai, NN;Blackmon, H;Goldfarb, J
在少精症的极端情况下，冷冻保存单个精子的方法将极大地使患者受益。本研究探讨了使用尼龙环冷冻保存少量精子，并描述了一种使用尼龙环冷冻单个精子的新技术。实验进行了比较精子恢复和生存能力后，冷冻保存在传统的小瓶和冷冻环。利用不同参数的废弃人类精子标本。该研究还检查了两种不同的甘油基冷冻保护剂。有和没有卵黄缓冲液。单精子冷冻保存时，选择5-10个精子，在显微镜和显微操作设备的辅助下，将其装入冷冻环。精子功能测试是在冷冻试管中冷冻的人和牛精子上进行的。冷冻试管中冷冻的微量精子表现出与冷冻试管中冷冻的对照样品相似的整体运动性和活力参数。单独选择冷冻保存在环上的精子容易恢复，解冻后运动能力一般较好。精子功能测试表明，冷冻保存在环上的人和牛精子注射到卵母细胞后都能进行精子头部去浓缩。冷冻环可能是一个极好的替代仓鼠带冷冻保存少量人类精子。
Methodology enabling cryopreservation of individual spermatozoa in extreme cases of oligozoospermia would tremendously benefit patients. This Study explores the use of a nylon loop for cryopreservation of small quantities of spermatozoa, and also describes a novel technique for freezing individual spermatozoa with this cryoloop. Experiments were conducted to compare sperm recovery and viability after cryopreservation in conventional vials versus on the cryoloops. Discarded human sperm specimens with varying parameters were utilized. The study also examines, two different glycerol-based cryoprotectants. with and without test yolk buffer. For single sperm cryopreservation, 5-10 spermatozoa were selected and loaded onto cryoloops with the aid of a microscope and micromanipulation equipment. Sperm function testing was performed on both human and bovine spermatozoa frozen on cryoloops. Microquantities of spermatozoa frozen on cryoloops exhibited overall motility and viability parameters similar to control samples frozen in cryovials. Individually selected spermatozoa cryopreserved on loops were easily recovered and post-thaw motility was generally good. Sperm function testing demonstrated that both human and bovine spermatozoa cryopreserved on loops were able to undergo sperm head decondensation when injected into oocytes. Cryoloops may be an excellent alternative to hamster zonae for cryopreserving small numbers of human spermatozoa.