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冷冻保护剂对家畜精液冷冻保存的研究进展 总被引：3，自引：0，他引：3
家畜精液冷冻保存可以实现在不同时间、不同地点的人工授精,能有效提高优良种公畜的利用率,扩大其基因传播范围,同时也是家畜遗传资源易位保存的主要方法之一。在精液冷冻保存过程中,选择合适的稀释液成分对解冻后精子品质的影响十分重要。精子对低温非常敏感,在稀释液中加入冷冻保护剂可有效提升精子解冻后活力。文章综述了近几年渗透性冷冻保护剂(甘油、乙二醇、二甲基甲酰胺、二甲基乙酰胺、二甲基亚砜)和非渗透性冷冻保护剂(卵黄、大豆卵磷脂、蔗糖、脱脂乳)在猪、牛、羊、鸡、兔以及骆驼上的应用与研究,旨在为进一步深入研究家畜精子冻融损伤机制提供理论依据。 相似文献
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牛精液冷冻与人工授精技术相结合，在牛品种改良、保护优良种质资源中发挥着重要作用。尽管冷冻后部分牛精子活力高达60%以上，但是冷冻后精子受损现象却普遍存在，尤其是受精能力与鲜精相比显著降低。作者详细介绍了冷冻-解冻过程对牛精子造成的主要损伤，包括精子的形态完整性、活力及遗传物质的改变等；阐述了冷冻造成精子损伤的主要原因，即细胞内冰晶形成和氧化应激反应及其产生的可能机制；并且对目前常用的提高冷冻精子质量的方法，如添加冷冻保护剂、优化冷冻程序以及添加抗氧化剂等进行了详细地综述；提出了在冷冻精液研究方面值得探索的问题，以期为家畜精液冷冻保存技术的进一步优化提供理论依据。 相似文献
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不同冷冻保护剂在鸡精液冷冻中的作用效果分析 总被引：3，自引：0，他引：3
本实验采用一定浓度的甘油、乙二醇、二甲基亚砜（ＤＭＳＯ）、二甲基乙酰胺（ＤＭＡ）作为冷冻保护剂，用含冷冻保护剂的稀释液将精液稀释后常温保存，观察精子活率，比较精子生存指数，并进行输精实验，发现在常温下对精子毒害作用最大的是甘油，其次是ＤＭＳＯ，而ＤＭＡ及乙二醇对精子的毒害作用最小。以一定浓度的４种冷冻保护剂将精液冷冻后观察解冻活率，发现以ＤＭＡ作为冷冻保护剂，解冻后精子活率最高。在输精实验中，以ＤＭＡ作为冷冻保护剂采用深阴道输精，取得了５０％的受精率。浅输精取得了４０％的受精率。 相似文献
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通过对不同浓度的甘油和乙二醇对牛精液冷冻保存效果的测定,探讨冷冻保护剂乙二醇与传统冷冻保护剂甘油的冷冻效果,并通过对葡萄糖和卵黄及Tris等成分的调整进一步分析了主要营养成分和缓冲体系对牛精液冷冻保存的影响程度。试验表明:1GL和EG对牛精子均有良好的冷冻保护能力,解冻后6h的活力GL组中以GL1.2和GL1.4的活力最高,均为0.50;EG组中以EG0.7的活力最高,达到0.53,高于0.35以上的国标。2其他营养成分的变化对GL和EG保护精子的效果有一定影响,特别是卵黄的作用比较明显;3在同等条件下,GL对精子的冷冻保护效果优于EG,EG组的活力衰竭快于GL组。 相似文献
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牛精液冷冻保存技术研究进展 总被引：1，自引：0，他引：1
《畜牧兽医杂志》2020,(2)
牛精液冷冻保存的应用促进了我国牛人工授精技术的发展,通过冷冻保存可以延长精液保存时间,扩大良种公牛精液的使用范围,提高种用价值,加速品种的育成和改良步伐。本文从牛冷冻精液稀释液的成分,卵黄稀释液与无动物源性稀释液的发展,冷冻保护剂对精子冷冻效果的影响,抗氧化剂和抗生素在牛精液冷冻保存中的应用等方面进行了综述,旨在为今后的牛精液冷冻保存研究工作提供一定的参考。 相似文献
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家畜精液的冷冻保存，是人工授精的一项重大改革，在精液冷冻技术发展过程中，由于稀释液中加入甘油作为保护剂，而使精液保存技术取得重大突破，但不同畜种的精子对甘油的反应不同。牛冷冻精液稀释液中甘油浓度或高或低对精子活率及受胎率关系不大，对猪和羊来说，甘油浓度增加时，冷冻精液的精子活率虽高，但受胎率极不理想。现将采用最新研制的无甘油糖类保护剂， 相似文献
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Use of Density Centrifugation for Delayed Cryopreservation of Stallion Sperm: Perform Sperm Selection Directly after Collection or after Storage? 
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下载免费PDF全文 A Heutelbeck H Oldenhof K Rohn G Martinsson JM Morrell H Sieme 《Reproduction in domestic animals》2015,50(1):76-83
Equipment for cryopreservation of stallion sperm is not always available. In such cases, diluted semen can be shipped to a facility for later cryopreservation. The aim of this study was to evaluate if selection of sperm via density centrifugation yields higher survival rates when cryopreservation is to be delayed (i.e. carried out after 1 day of storage at 5°C). Two‐layer iodixanol as well as single‐layer Androcoll density centrifugation were tested and compared with samples prepared with standard centrifugation. Special emphasis was placed on comparing centrifugation on the day of semen collection with centrifugation after 1‐day refrigerated storage. Sperm morphology and motility as well as membrane and chromatin integrity were evaluated before and after centrifugation. Sperm motility and membrane integrity were also assessed after cryopreservation. It was found that both two‐ and single‐layer density centrifugation processing resulted in higher percentages of morphologically normal and motile sperm with higher membrane and chromatin integrity, as compared to standard centrifugation or diluted samples. Differences were only in the order of magnitude of 5%. Recovery rates after density centrifugation were only approximately 30–40%. When cryopreservation was carried out after 1‐day refrigerated storage, centrifugation processing of sperm directly after semen collection resulted in higher percentages of plasma membrane intact sperm post‐thaw as compared to performing centrifugation processing of stored sperm just prior to cryopreservation. No significant differences in progressively motile sperm post‐thaw were seen. Taken together, for delayed cryopreservation, it is best to perform density centrifugation directly after collection rather than immediately prior to cryopreservation. 相似文献
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采用5% 二甲乙酰胺(DMA)(V/V)完全替代甘油,比较乳糖、海藻糖对精液冷冻保存效果的影响。结果表明,海藻糖显著提高了冷冻——解冻后精子成活力(49.32%±1.52%)与顶体完整性 (47.33%±1.16%)(P<0.05)。然后利用海藻糖替代乳糖,评价不同浓度的DMA对公猪精液冷冻保存的影响。结果表明,当DMA添加量为4%时,解冻后精子活率、成活力、顶体完整率分别为(45.17±0.56)%、(50.33±0.67)%、(48.30±1.44)%,均显著高于3% DMA、6% DMA添加组(P<0.05),精子活率显著高于5% DMA添加组(P<0.05),但精子成活力、顶体完整性与其差异不显著(P>0.05)。因此,当利用海藻糖作为冷冻保存基础稀释液,DMA最适添加量为4%。 相似文献
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Damage occurring to spermatozoa during cryopreservation results in a loss of motile cells and cells that are functionally normal, compared with fresh sperm samples. Treating bull sperm with cholesterol-loaded cyclodextrins (CLC) before cryopreservation results in increased sperm cryosurvival. However, in previous studies, CLC were always added to sperm samples that had been highly diluted. The aim of this study was to develop a procedure for adding CLC to whole bull ejaculates that would optimize sperm cryosurvival. Adding 2 or 4 mg of CLC/120 x 10(6) sperm to sperm samples ranging in concentration from 120 to 2,000 x 10(6) sperm/mL resulted in greater (17 to 28 percentage points; P < 05) numbers of live cells compared with control samples (no CLC treatment), regardless of the sperm concentration, except for samples at 120 x 10(6) sperm/mL treated with 4 mg of CLC. Incubating sperm with CLC at 23 or 37 degrees C before cryopreservation resulted in similar sperm cryosurvival. The cooling rate used to cryopreserve CLC-treated cells did not affect sperm cryosurvival. Finally, adding CLC to undiluted ejaculates (2 mg of CLC/120 x 10(6) sperm) resulted in greater percentages of live sperm compared with the control samples (62 vs. 45%; P < 0.05), although the percentages of motile sperm were similar for both CLC-treated and control samples (58%). In conclusion, bull sperm cryo-survival can be improved if spermatozoa are treated with CLC before freezing. In addition, CLC can be added to fresh ejaculates at either 23 or 37 degrees C. This technique is simple, practical, and can be easily integrated into current cryopreservation protocols. 相似文献
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This study investigated the apoptosis-like events associated with cryopreservation process and their relationship with cryocapacitation in buffalo (Bubalus bubalis) sperm. A total of 49 semen ejaculates from seven bulls were studied for structural changes in sperm following cryopreservation. Apoptotic changes were detected by assays specific for translocation of phosphatidylserine (PS) to the cell surface, alterations in membrane permeability and mitochondrial membrane potential (MMP), and DNA integrity. A significant (p < 0.01) percentage of cryopreserved sperm showed externalization of PS and early apoptotic changes and lowered MMP when compared with the fresh sperm. Freezing and thawing of sperm increased permeability to YOPRO-1, an impermeant fluorescent dye. However, on TUNEL staining, cryopreserved sperm showed no breach in DNA integrity. The sperm capacitation status was evaluated by chlortetracycline (CTC) fluorescence pattern, in which a significant (p < 0.01) percentage of cryopreserved sperm were found to be capacitated. The capacitated sperm (Pattern B) was positively correlated with the aforementioned apoptotic events. In conclusion, cryopreservation process induced early apoptosis-like changes in buffalo sperm, and a close link exists between cryocapacitation and apoptosis during cryopreservation of sperm. 相似文献
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MC Esteso MR Fernández-Santos AJ Soler V Montoro A Quintero-Moreno JJ Garde 《Reproduction in domestic animals》2006,41(3):241-246
Computer-automated sperm-head morphometry was used in this study to determine the effects of cryopreservation on red deer sperm-head morphometry. Epididymal sperm samples were collected from 40 mature stags and were divided. One portion was diluted at room temperature in a Tris-citrate egg yolk medium, containing 6% glycerol. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours. After thawing, sperm smears were prepared as described above. All slides were air dried and stained with Hemacolor. The sperm-head dimensions for length, width, area, perimeter and shape factor (length/width), for a minimum of 135 spermatozoa were determined for each slide by means of the Sperm-Class Analyser (SCA). Firstly, our results show that cryopreservation substantially reduced (p < 0.001) sperm motility and plasma membrane and acrosome integrities. In addition, sperm heads were significantly smaller in cryopreserved spermatozoa than in the companion extended samples for area (32.05 microm2 vs 32.56 microm2; p < 0.05), length (8.46 microm vs 8.53 microm; p < 0.0001) and shape factor (1.833 vs 1.849; p < 0.0001) for all stags. These differences were found within 29 of 40 stags (75%) for at least three of the morphometric parameters. The individual variability (CV) of sperm head measurements from extended samples was negatively correlated (p < 0.005) with the per cent of change in sperm head measurements after cryopreservation for area (r = -0.465), width (r = -0.483) and perimeter (r = -0.375). Thus, the lower the sperm head variability in the extended samples, the greater the sperm change as a consequence of the cryopreservation. These results suggest that the variability (heterogeneity) in sperm head dimensions of individual stags may be a good indicator of sperm freezability. 相似文献
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Yang S Ping S Ji S Lu Y Niu Y Wang H Ji W Si W 《The Journal of reproduction and development》2011,57(6):737-743
The objective was to examine the effect of seminal plasma on cryopreservation of sperm from rhesus macaques. Sperm cryosurvival was evaluated by sperm motility and acrosomal integrity. Compared with slow cooling (-0.4 C/min) from 37 C (body temperature) to 4 C, rapid cooling (-16 C/min) caused cold shock in rhesus macaque sperm. The cryosurvival of sperm was decreased regardless of the presence or absence of seminal plasma (P<0.05). However, the presence of seminal plasma during cold shock at a rapid cooling rate improved sperm motility and acrosomal integrity in individual monkeys. Male-to-male variation in sperm cryosurvival was observed after cryopreservation (P<0.05), and the presence of seminal plasma during sperm cryopreservation improved sperm motility and acrosomal integrity in individual monkeys (P<0.05). Furthermore, by adding seminal plasma from monkeys with good sperm cryosurvival to sperm freezing extender, the frozen-thawed motility and acrosomal integrity of sperm from monkey with poor cryosurvival were improved (P<0.05). The present study indicated that seminal fluid is beneficial to sperm undergoing cold shock or cryopreservation in individual monkeys. The cryosurvival of sperm from rhesus macaques with poor sperm freezability could be improved by the presence of seminal plasma from males with good sperm cryosurvival. This finding provides a useful method for genetic preservation in this important species. 相似文献
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通过对精液冷冻保存的细胞反应原理的阐述 ,指出冷冻保护剂甘油对精液保存具有利弊效应 ,提出精子膜脂组成的差异使得不同品种的精子对冷冻损伤的易感性不同。雌性生殖道解剖结构的品种差异 ,精子形态 ,精子运行机制的细微差异 ,人工授精时间及精子的运行能力 ,采精方式等因素对精液冷冻保存和人工授精的成功有决定性作用。研究精子质膜的生物学特性可解决低活力精子的问题 ,然而这并不能解决冷冻后精子质量的个体差异。对精细胞基因组的研究可以找出这些个体的遗传差异。因此 ,冷冻精子和精原细胞 (用于细胞外注射 )的差异已经成为完整基因组问题。 相似文献
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精子冷冻是一种简单、经济、高效的生物遗传资源保存方法,目前已经应用到许多领域,并成为治疗人类不育、牲畜繁殖和生物资源多样性保护的重要手段。在精子冷冻保存过程中的适当阶段加入防冻剂,能够有效的避免或减少精子的冷冻损伤和寒害,从而提高冷冻—解冻精子的活力。作者就防冻剂的种类、防冻剂的选择及加入与去除的方式对冷冻—解冻哺乳动物精子活力的影响作一简要综述。 相似文献
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Kim S Lee YJ Ji DB Kim YJ 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(7):961-963
This study was performed to evaluate the use of dimethylacetamide (DMA) and dimethyl sulfoxide (DMSO) in boar sperm cryopreservation. Semen from eight boars was cryopreserved following treatment with 3, 5, and 7% DMA and DMSO, and 3% glycerol (control). After thawing, sperm conventional parameters and membrane integrities were evaluated. There were no significant differences among different DMA concentrations in all evaluations. Membrane intactness were higher in 5% and 7% DMSO than 3% DMSO (P<0.05). Sperm motility of 5% DMSO was lower than that of 3% glycerol (P<0.005), and membrane intactness were lower in 5% DMA and DMSO than 3% glycerol (P<0.05). DMA and DMSO didn't improve sperm quality and glycerol remains the most useful for boar sperm cryopreservation. 相似文献