Zavos P M, Sofikitis N, Toda T, Miyagawa I
Department of Animal Sciences, University of Kentucky, Lexington 40546.
Tohoku J Exp Med. 1991 Dec;165(4):283-90. doi: 10.1620/tjem.165.283.
Motile, morphologically normal human spermatozoa can be separated from semen via a disposable SpermPrep filtration method. This method was employed with great success using frozen-thawed spermatozoa. The present study examined the qualitative characteristics of filtered spermatozoa via a new SpermPrep II filtration method in conjunction with the short-term freeze preservation of these spermatozoa. The isolation procedure yielded populations of spermatozoa with very high percentage motility and progressive motility grade (0-4) and which were free of seminal debris. For 20 semen samples the prefreeze values of percentage of motility and grade for the fresh and post-filtered were 58.6% and 3.1 vs 84.1% and 3.6, respectively. The sperm freeze preservation procedure involved dilution of the spermatozoa in Test-Yolk buffer (TYB) with 7% glycerol (v/v) and freezing and thawing in a conventional manner. Post-thaw percent motility, and survival % were substantially higher (p less than 0.05) for the filtered fraction than for the parent semen. The filtered fractions yielded post-thaw mean values of motility percent and grade of 68.9 and 3.4 vs 47.6 and 3.0 for the parent semen, respectively. Survival following incubation at 37 degrees C for 3 hours yielded respective values of 75.9% and 53.2% (p less than 0.05). It was concluded that the major factor in improving post-thaw quality recovery overall was the filtration via the SpermPrep II as applied in this study. This technique could have significant clinical applications in the use of frozen-thawed specimens for non-coital reproduction purposes.
可通过一次性精子制备过滤法从精液中分离出具有运动能力、形态正常的人类精子。该方法在使用冻融精子时取得了巨大成功。本研究通过一种新的精子制备II过滤法结合这些精子的短期冷冻保存，研究了过滤后精子的质量特征。分离程序产生了具有非常高的运动百分比和渐进运动等级（0 - 4）且无精液碎片的精子群体。对于20个精液样本，新鲜和过滤后精子的运动百分比和等级的冻前值分别为58.6%和3.1，以及84.1%和3.6。精子冷冻保存程序包括在含有7%甘油（v/v）的测试卵黄缓冲液（TYB）中稀释精子，并以常规方式冷冻和解冻。解冻后，过滤部分的运动百分比和存活率显著更高（p小于0.05），高于原始精液。过滤部分解冻后的运动百分比平均值和等级分别为68.9和3.4，而原始精液分别为47.6和3.0。在37摄氏度孵育3小时后的存活率分别为75.9%和53.2%（p小于0.05）。得出的结论是，总体上提高解冻后质量恢复的主要因素是本研究中应用的通过精子制备II进行的过滤。该技术在将冻融样本用于非性交生殖目的方面可能具有重要的临床应用。
