Sieme Harald, Oldenhof Harriëtte
Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559, Hannover, Germany,
Methods Mol Biol. 2015;1257:343-52. doi: 10.1007/978-1-4939-2193-5_16.
Fertility rates with artificial insemination are highest with good-quality sperm samples. Therefore, nonviable sperm, cellular debris, and seminal plasma are preferably removed from semen samples prior to use or for preservation. Such compounds are sources where reactive oxygen species are generated during storage or upon cryopreservation, impairing sperm function. In this chapter we describe methods to remove seminal plasma and cellular debris from sperm samples, and for selecting morphologically normal motile sperm. The methods that are described here include: ordinary centrifugation, sperm swim-up, glass wool and Sephadex filtration/adherence, and single-layer as well as discontinuous two-layer iodixanol density gradient centrifugation.
人工授精时,使用优质精子样本的受精率最高。因此,在使用或保存精液样本之前,最好去除其中无活力的精子、细胞碎片和精浆。这些化合物是储存或冷冻保存过程中产生活性氧的来源,会损害精子功能。在本章中,我们描述了从精子样本中去除精浆和细胞碎片以及选择形态正常、有活力精子的方法。这里描述的方法包括:普通离心、精子上游法、玻璃棉和葡聚糖凝胶过滤/黏附法,以及单层和不连续双层碘克沙醇密度梯度离心法。