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使用流式细胞术和计算机辅助精子分析仪评估家猫(Felis catus s. domestica,L. 1758)附睾精子的选定质量参数。

Assessment of selected quality parameters of epididymal cat (Felis catus s. domestica, L. 1758) sperm using flow cytometry method and computer assisted sperm analyser.

作者信息

Siemieniuch M J, Woclawek-Potocka I

机构信息

Department of Reproductive Immunology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland.

出版信息

Reprod Domest Anim. 2008 Oct;43(5):633-7. doi: 10.1111/j.1439-0531.2007.00958.x. Epub 2008 Mar 23.

Abstract

The improvement of biotechnical methods connected with fast and precise semen quality assessment and its utilization in assisted reproductive techniques is an urgent necessity in felids. The aim of this study was to evaluate some quality parameters (i.e. the viability and share of cells with intact plasma membrane) of epididymal sperm of cats using the flow cytometry method and computer-assisted sperm analysis (CASA) examination. The material consisted of epididymal spermatozoa flushed from 22 pairs of epididymes after routine neutering procedures obtained from domestic cats aged between 8 and 36 months. The epididymes were cut and incubated with an extender without egg yolk. The samples were assessed for sperm viability (Live/Dead Sperm Viability Kit), percentage of subtle membrane changes (Apoptosis Detection Kit) and motility using FACScalibur flow cytometer and assisted sperm analyser HTM IVOS version 12.2. The flow cytometry method revealed 71.3% and 84.4% of live sperm using Live/Dead Sperm Viability Kit and Apoptosis Detection Kit respectively. The population of early-apoptotic and late-apoptotic sperm were 0.8% and 1.1% respectively. The CASA examination found 51.5% of motile sperm. However, the motility examination under light microscope revealed 69.5% of motile sperm. The data revealed an indistinctive per cent of apoptotic cells and 18.9% and 15.6% of dead cells using Live/Dead Sperm Viability Kit and Apoptosis Detection Kit, respectively, which indicate that the sperm obtained after flushing the epididymis possess potential properties for further assisted reproduction techniques.

摘要

改进与快速精确的精液质量评估相关的生物技术方法及其在猫科动物辅助生殖技术中的应用是当务之急。本研究的目的是使用流式细胞术和计算机辅助精子分析(CASA)检测评估猫附睾精子的一些质量参数(即具有完整质膜的细胞的活力和比例)。材料包括从22对附睾中冲洗出的附睾精子,这些附睾是在对8至36个月大的家猫进行常规绝育手术后获得的。将附睾切开并与不含蛋黄的稀释液一起孵育。使用FACScalibur流式细胞仪和辅助精子分析仪HTM IVOS版本12.2评估样品的精子活力(活/死精子活力试剂盒)、细微膜变化百分比(凋亡检测试剂盒)和运动能力。流式细胞术方法使用活/死精子活力试剂盒和凋亡检测试剂盒分别显示活精子的比例为71.3%和84.4%。早期凋亡精子和晚期凋亡精子的比例分别为0.8%和1.1%。CASA检测发现51.5%的精子具有运动能力。然而,在光学显微镜下进行的运动能力检测显示69.5%的精子具有运动能力。数据显示,使用活/死精子活力试剂盒和凋亡检测试剂盒时,凋亡细胞的百分比不明显,死细胞的比例分别为18.9%和15.6%,这表明冲洗附睾后获得的精子具有用于进一步辅助生殖技术的潜在特性。

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