Anders Johannisson , Jane M. Morrell , Margareta Wallgren
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引用次数: 0
Abstract
Not all boar sperm samples survive cryopreservation well. A method of eliminating damaged sperm might enable more cryopreserved boar semen to be used for pig breeding. In this study we investigated the use of Magnetic Activated Cell sorting (MACS) to eliminate damaged sperm from thawed boar semen samples. The thawed samples were mixed with Dead cell removal particles and were applied to the column in a SuperMACS II. Different fractions were collected: Original sample (O), Flow-through (FT), and Eluate (E). Sperm membrane integrity, mitochondrial membrane potential and reactive oxygen species were evaluated by flow cytometry after staining with SYBR 14 and propidium iodide, or 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolylcarbocyanine iodide, or hydroethidine and dichlorodihydrofluorescein diacetate, respectively. The FT samples had increased membrane integrity, a greater proportion of sperm with high mitochondrial membrane potential and a greater proportion of sperm negative for hydrogen peroxide than O samples (P<0.0001), which in turn had increased membrane integrity than E samples (P <0.0001). However, differences were seen between boars. The FT samples had increased values of live, superoxide positive sperm than O samples (P <0.0001) and O samples had greater values than E samples (P <0.0001), while there was no effect of boar. Sperm quality was best in the FT fraction, comprising approximately 32% of the sperm sample. In conclusion, although there were differences between boars, MACS separation can improve sperm quality in thawed semen samples. It would be interesting to see if this improvement is reflected in fertility outcomes.
并非所有公猪精子样本都能很好地冷冻保存。一种消除受损精子的方法可使更多冷冻保存的公猪精液用于种猪繁殖。在这项研究中，我们研究了使用磁激活细胞分拣技术（MACS）消除解冻公猪精液样本中受损精子的方法。解冻后的样本与去死皮细胞颗粒混合后，进入超级磁性细胞分拣系统 II 的色谱柱。收集不同的馏分：原始样品（O）、流出液（FT）和洗脱液（E）。分别用 SYBR 14 和碘化丙啶，或 5′、6、6′-四氯-1、1′、3、3′-四乙基苯并咪唑羰花青碘化物，或氢化乙锭碱和二氯二氢荧光素二乙酸酯染色后，用流式细胞仪评估精子膜完整性、线粒体膜电位和活性氧。与 O 样品相比，FT 样品的膜完整性增加，线粒体膜电位高的精子比例增加，过氧化氢阴性的精子比例增加（P<0.0001），而 O 样品的膜完整性又比 E 样品增加（P<0.0001）。然而，公猪之间也存在差异。FT 样品的活精子、超氧化物阳性精子的数值比 O 样品高（P <0.0001），O 样品的数值比 E 样品高（P <0.0001），而公猪没有影响。FT部分的精子质量最好，约占精子样本的32%。总之，尽管公猪之间存在差异，但 MACS 分离能提高解冻精液样本的精子质量。我们有兴趣了解这种改善是否反映在生育结果上。
期刊介绍：
Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction.
The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques.
The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.
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