通过Sephadex柱过滤,选择高质量的冷冻解冻牛精子亚群。黄体酮的功能作用评价

L.M. Vincenti, M. Fiol de Cuneo , R.D. Ruiz , A.C. Martini, A.A. Ponce
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引用次数: 3

摘要

低温保存对哺乳动物精子的功能特性有不利影响,这是公认的。处理冻融配子的传统方法,如离心,会产生额外的损害。在本工作中,我们研究了用Sephadex柱过滤(SF组)或用离心清洗(100 g, 10 min, 2次)(W组)处理牛冷冻精液的效果;将两种方法获得的结果与未处理的样品(C组)进行比较。研究了体外添加10 μM、20 min的黄体酮对精子功能活性的影响。评估的精子参数包括浓度、活力(进行性或非进行性细胞)、活力和顶体反应。在时间0(处理后立即)或在使能条件下孵育4小时后测量它们。精子浓度(× 10−6):C组37.5±5.4,W组8.3±2.1,SF组12.5±2.9。SF组的运动百分率、进展百分率、活体细胞百分率和顶体完整配子百分率均显著高于W组和c组。SF组在使能条件下孵育4 h后,孕酮显著增加顶体反应细胞的数量,而在不含肝素的条件下孵育时,这一参数没有改变。运动能力和生存能力不受激素的影响。我们的结论是,Sephadex过滤法是一种足够的工具,以获得高质量的精子亚群,通过运动性,活力和顶体完整性来评估;此外,我们的结果有力地支持,黄体酮将是一个生理诱导性的顶体反应,因为在其他物种中,牛。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Selection of a high quality subpopulation of frozen thawed bovine spermatozoa by filtration in a Sephadex column. An assessment of functional effects of progesterone

It is accepted that cryopreservation exerts deleterious effects on functional characteristics of mammalian spermatozoa. Conventional procedures for processing frozen-thawed gametes, such as centrifugation, produce additional damage. In the present work, we investigated the efficacy of processing bovine cryopreserved semen by filtration in a Sephadex column (SF group) or by washing by centrifugation (100 g, 10 min, twice) (W group); the results obtained from both procedures were compared to untreated samples (C group). The effects of in vitro addition of progesterone (10 μM, 20 min) upon sperm functional activity were studied also. The evaluated sperm parameters were concentration, motility (progressive or non progressive cells), viability and acrosome reaction. They were measured at time 0 (immediately after processing) or after 4 h incubation in capacitating conditions. Sperm concentration was (× 10−6): 37.5 ± 5.4 in C, 8.3 ± 2.1 in W and 12.5 ± 2.9 in SF. The percentages of motile, progressive, viable or acrosome intact gametes were significantly higher in SF than in W or in C. in SF group, after 4 h incubation in capacitating conditions, progesterone increased significantly the population of acrosome reacted cells whereas this parameter was not modified when the cells were incubated in absence of heparin. Motility and viability were not modified by the hormone. We conclude that Sephadex filtration method is an adequate tool to obtain a subpopulation of spermatozoa with superior quality, as assessed by motility, viability and acrosomal integrity; besides, our results strongly support that, as in other species, progesterone would be a physiological inductor of acrosome reaction in bovine.

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