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BoviPure® 密度梯度离心程序可提高新鲜和冷冻保存的犬附睾精子质量。

BoviPure® density-gradient centrifugation procedure enhances the quality of fresh and cryopreserved dog epididymal spermatozoa.

机构信息

Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, EC010205 Cuenca, Ecuador.

Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, EC010205 Cuenca, Ecuador.

出版信息

Anim Reprod Sci. 2022 Jul;242:107003. doi: 10.1016/j.anireprosci.2022.107003. Epub 2022 May 19.

DOI:10.1016/j.anireprosci.2022.107003
PMID:35653822
Abstract

BoviPure® is a salt solution containing colloidal silica particles coated with silane used to select sperm (e.g., ruminants) by density-gradient centrifugation (DGC). This research assessed the suitability of the BoviPure-DGC and swim-up methods for selecting dog epididymal sperm in fresh, chilled and frozen-thawed samples on post-treatment sperm quality. Sperm samples (n = 60 epididymides) were recovered by retrograde flushing from thirty orchiectomized adult dogs. Thereafter, 20 sperm pools, containing sperm aliquots of three randomly selected animals, were used for chilling (at 5 ºC for 24 h) and freezing (in liquid nitrogen vapors). Sperm selection by BoviPure-DCG and swim-up was performed in both individual and pooled samples, including non-selected samples as controls. Overall, after BoviPure-DGC selection a higher sperm retrieval rate was obtained than the swim-up selection in both individual (P < 0.05) and pooled (P < 0.01) samples. BoviPure-DGC improved (P < 0.05) the total (TM) and progressive (PSM) sperm motilities, curvilinear (VCL) and straight-line (VSL) velocities, linearity (LIN), wobble (WOB), beat-cross frequency (BCF), and integrity of plasmatic (IPM) and acrosomal (IAM) membranes of individual samples in comparison with non-selected samples. In pooled samples, however, the BoviPure-DGC improved (P < 0.05) the PSM, VCL, WOB, and IPM of chilled and frozen-thawed samples. The swim-up method improved (P < 0.05) only some kinematic variables of the individual (VCL, WOB and BCF) and cryopreserved pooled samples (VCL and ALH) in comparison with non-selected samples. In conclusion, BoviPure-DGC was more effective for recovering and selecting both fresh and cryopreserved dog epididymal sperm than the swim-up procedure improving the kinematic variables, and membranes intactness.

摘要

BoviPure® 是一种含有胶体硅颗粒的盐溶液,表面涂有硅烷,用于通过密度梯度离心(DGC)选择精子(例如反刍动物)。本研究评估了 BoviPure-DGC 和泳动法在新鲜、冷藏和冷冻解冻样本中选择狗附睾精子的适用性,研究对象为处理后的精子质量。通过逆行冲洗从 30 只去势成年犬的睾丸中回收精子样本(n=60 个附睾)。此后,使用 20 个精子池,每个精子池包含 3 只随机选择的动物的精子等分试样,进行冷藏(5°C 24 小时)和冷冻(在液氮蒸气中)。在个体和混合样本中都进行了 BoviPure-DCG 和泳动法的精子选择,非选择样本作为对照。总体而言,BoviPure-DGC 选择后的精子回收率高于泳动法选择,无论是在个体样本(P<0.05)还是混合样本(P<0.01)中。与非选择样本相比,BoviPure-DGC 提高了个体样本的总(TM)和渐进(PSM)精子活力、曲线(VCL)和直线(VSL)速度、直线性(LIN)、摆动(WOB)、鞭打交叉频率(BCF)以及质膜(IPM)和顶体膜(IAM)的完整性(P<0.05)。然而,在混合样本中,BoviPure-DGC 仅提高了冷藏和冷冻解冻样本的 PSM、VCL、WOB 和 IPM(P<0.05)。与非选择样本相比,泳动法仅提高了个体样本(VCL、WOB 和 BCF)和冷冻保存的混合样本(VCL 和 ALH)的一些运动学变量(P<0.05)。总之,与泳动法相比,BoviPure-DGC 更有效地回收和选择新鲜和冷冻保存的狗附睾精子,提高了运动学变量和膜的完整性。

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