聚乙烯醇和聚乙烯吡咯烷酮制备解冻的猪精ICSI受精用精子

O. J. Lyzohub
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摘要

本研究旨在探讨PVP(聚乙烯吡咯烷酮)和PVA(聚乙烯醇)培养基对公猪脱存精液的影响及其制备方法,以优化人工授精的生物技术途径。这项研究使用了Myrhorod品种Dnipro 641的一头公猪的冷冻精子。遗传物质在以M.V. Zubets NAAS命名的动物遗传资源银行IABG中保存了8年。精子悬浮液在+37℃水浴中解冻5分钟至完全解冻。在Sp-TALP培养基中,用向上游动法从冷冻剂和稀释液中分离精子。精子在10.0% PVP溶液中孵育10 min后,活动力下降68.2% (P < 0.05),为3.4%;孵育10 min后,活动力下降至1.4% (P <0.01),为初始活动力的10倍。10.0%的PVA孵育10 min后活动性下降了37.4% (P <0.05),达到6.7%,10 min后活动性下降到5.7% (P < 0.01),比初始活动性降低了1.8倍。结果表明,在PVP浓度为10.0%的情况下,未保存的猪精子的活力比初始水平下降了86.9% (P <0.01),这使得无法通过胞浆内单精子注射(ICSI)选择合适的精子。结果表明,10.0% PVA溶液可使猪精子的初始活动力降低46.7% (P <0.01)。结果表明,在5.0% PVA溶液中孵育未保存的猪精时,活动性仅比初始降低28.0% (P <0.05),当使用冷冻保存的猪精时,活动性是最佳的,这种材料对操作者来说是有限的,方便的,对卵母细胞是安全的。
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Usage of polyvinylalcohol and polyvinylpyrrolidone for preparion of thawed ejaculated boar sperm for fertilization by ICSI
The aim of the study was to explore the effect of PVP (polyvinylpyrrolidone) and PVA (polyvinyl alcohol) media on deconserved ejaculated boar sperm and their preparation for artificial insemination to optimize biotechnological approaches. The studies used ejaculated cryopreserved sperm of a boar of the Myrhorod breed Dnipro 641. Genetic material was stored in the Bank of Genetic Resources of Animals IABG nnamed after M.V. Zubets NAAS for eight years. The sperm suspension was thawed in a water bath at +37 °C for 5 min until completely thawed. Separation of sperm from cryopreservative agent and diluent was performed using the swim up method in Sp-TALP medium. After the presence of sperm in the 10.0% solution of PVP for 10 min, motility decreased by 68.2% (P < 0.05) and amounted to 3.4%, and after the next 10 min of incubation decreased to 1.4% (P <0.01), which is 10 times lower than the initial mobility. In 10.0% of PVA mobility after 10 min of incubation decreased by 37.4% (P <0.05) and amounted to 6.7%, and after 10 min decreased to 5.7% (P < 0.01), which is 1.8 times lower than the initial mobility. It was found that in the case of 10.0% of PVP solution ejaculated deconserved boar sperm lose motility by 86.9% (P <0.01) from the initial motility, which makes it impossible to select a suitable sperm for fertilization by ICSI (intracytoplasmic sperm injection). It is shown that 10.0% PVA solution can be used for immobilization of boar sperm, as it reduces motility by 46.7% (P <0.01) of the initial sperm motility. It is proved that the mobility in the case of incubation of deconserved ejaculated boar sperm in 5.0% PVA solution decreases only by 28.0% (P <0.05) from the initial, which is optimal when using cryopreserved boar sperm, material which are limited and convenient for the operator and safe for oocytes.
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