Zavos P M, Correa J R, Sofikitis N, Kofinas G D, Zarmakoupis P N
Andrology Institute of Lexington, University of Kentucky, Lexington 40546, USA.
Tohoku J Exp Med. 1995 Jun;176(2):75-81. doi: 10.1620/tjem.176.75.
The objective of this study was to determine if spermatozoa, following short-term cryostorage at 5 degrees C in Test-Yolk buffer (TYB; ZBL, Inc., Lexington, KY, USA), could be recovered and improved via the SpermPrep filtration method and to assess the possibly enhanced fertilizing capacity of the selected spermatozoa. Semen specimens from 20 men were collected, evaluated, diluted 1:1 (v/v) with TYB, divided into aliquots and cooled to 5 degrees C for 24 and 48 hr. Semen samples were assessed for volume, sperm count, percentage and grade of motility, percentage of morphologically normal spermatozoa and outcome of the sperm penetration assay (SPA). After storage, aliquots were rewarmed at 37 degrees C, centrifuged, and the pellet was resuspended in 1.0 ml of SpermPrep media (ZBL, Inc.). Following 15 min of incubation, the rewarmed spermatozoa were filtered via the SpermPrep I filtration column (ZBL, Inc.) and assessed accordingly. The results obtained in this study indicate that the short-term cryostorage procedure yielded spermatozoa of adequate qualitative characteristics when compared to the fresh spermatozoa. Furthermore, filtration of rewarmed specimens yielded spermatozoa of significantly higher qualitative characteristics and superior fertilizing capacity following a short-term cryostorage period in TYB when compared to fresh and rewarmed spermatozoa (p < 0.05). This method of short-term cryostorage in TYB and selection of superior spermatozoa via the SpermPrep filtration method could further enhance the fertilizing ability of patients who produce spermatozoa characterized by deficient capacitation, acrosome reaction and subsequent fertilization.
本研究的目的是确定精子在5℃下于测试-蛋黄缓冲液（TYB；ZBL公司，美国肯塔基州列克星敦）中短期冷冻保存后，是否可通过精子制备过滤方法恢复并得到改善，并评估所选精子可能增强的受精能力。收集了20名男性的精液标本，进行评估后，用TYB按1:1（v/v）稀释，分成小份并冷却至5℃保存24小时和48小时。对精液样本进行体积、精子计数、活力百分比和等级、形态正常精子百分比以及精子穿透试验（SPA）结果的评估。保存后，将小份样本在37℃复温，离心，沉淀重悬于1.0 ml精子制备培养基（ZBL公司）中。孵育15分钟后，将复温的精子通过精子制备I过滤柱（ZBL公司）过滤并进行相应评估。本研究获得的结果表明，与新鲜精子相比，短期冷冻保存程序产生的精子具有足够的质量特征。此外，与新鲜和复温的精子相比，在TYB中短期冷冻保存后，复温样本的过滤产生了质量特征显著更高且受精能力更强的精子（p < 0.05）。这种在TYB中短期冷冻保存并通过精子制备过滤方法选择优质精子的方法，可进一步提高产生具有获能、顶体反应及后续受精缺陷特征精子的患者的受精能力。
