Cryopreservation of domestic animal sperm cells
Cryopreservation of domestic animal sperm cells
复制标题
DOI：
10.1007/s10561-008-9081-4
复制
发表时间：
2009-02-01
影响因子：
1.5
通讯作者：
Mascarenhas, R. D.
中科院分区：
文献类型：
--
作者：
Barbas, J. P.;Mascarenhas, R. D.
精子细胞是男性精子发生的终点，具有特殊的解剖和代谢特征。精子冷冻保存和储存目前需要液氮或超低冷冻方法进行长期或短期储存，这需要日常维护和广泛的空间要求。精子保存具有人工生殖技术、物种保护和临床医学等多种用途。储存温度、冷却速度、延长剂的化学成分、冷冻保护剂浓度、活性氧（ROS）、精浆成分和卫生控制等因素的组合是影响精子寿命的关键因素。精子保存方案因动物物种的固有特殊性而有所不同，这些特殊性改变了用于冷藏和冷冻的扩展剂。冷冻精子细胞的填充剂含有缓冲液、碳水化合物（葡萄糖、乳糖、棉子糖、蔗糖和海藻糖）、盐（柠檬酸钠、柠檬酸）、蛋黄和抗生素。我们的实验室正在研究使用不同的冷冻保护剂，如海藻糖或甘油，以及精液填充剂中不同浓度的蛋黄和其他成分。已经测试了几种冷却速率来冷冻精子细胞。使用更快的冷却速度（15-60摄氏度/分钟）可以提高精子在冻融后的最佳存活率，但需要更多的研究来为每种动物找到适当的冷却速度。一些本地品种的公绵羊和山羊正在被用于在EZN进行的研究。这些雄性的精液已被冷冻并作为葡萄牙动物种质库的一部分储存起来。在小型反刍动物中，已经观察到冷冻精液质量的个体差异，这表明精子对冷冻方法的易感性存在特定差异，在山羊雄性中尤其明显。来自小反刍动物的优质冷冻精液被用于宫颈人工授精研究，旨在提高选定畜群的生产参数。
Sperm cells are the endpoint of male spermatogenesis and have particular anatomic and metabolic features. Sperm cryopreservation and storage currently require liquid nitrogen or ultralow refrigeration methods for long or short term storage, which requires routine maintenance and extensive space requirements. Conserving sperms have several purposes such as artificial reproductive technologies (ART), species conservation and clinical medicine. The combinations of storage temperature, cooling rate, chemical composition of the extender, cryoprotectant concentration, reactive oxygen species (ROS), seminal plasma composition and hygienic control are the key factors that affect the life-span of spermatozoa. Sperm preservation protocols vary among animal species owing to their inherent particularities that change extenders used for refrigeration and freezing. Extenders for freezing sperm cells contain buffers, carbohydrates (glucose, lactose, raffinose, saccharose and trehalose), salts (sodium citrate, citric acid), egg yolk and antibiotics. The use of different cryoprotectants, like trehalose or glycerol, as well as different concentrations of egg yolk and other constituents in semen extenders are being studied in our laboratory. Several cooling rates have been tested to freeze sperm cells. The use of faster rates (15-60A degrees C/min) gives rise to best sperm survivals after freezing-thawing, but more studies are needed to find the adequate cooling rates for each animal species. Sheep and goat males of some native breeds are being used in studies performed in EZN. Semen from those males has been frozen and stored as part of the Portuguese Animal Germplasm Bank. In small ruminants, individual variations in the quality of frozen semen have been observed, suggesting specific differences in sperm susceptibility to freezing methods, particularly obvious in goat males. Best quality frozen semen from small ruminants is being used in cervical artificial insemination studies aiming to increase productive parameters in selected flocks.