Risopatrón J, Catalán S, Miska W, Schill W-B, Sánchez R
Faculty of Medicine, Universidad de La Frontera, Temuco, Chile.
Reprod Domest Anim. 2002 Dec;37(6):347-51. doi: 10.1046/j.1439-0531.2002.t01-1-00380.x.
Sperm culture media used for in vitro fertilization (IVF) procedures are important factors concerning the viability, motility and acrosomal integrity of spermatozoa. The aim of this study was to investigate the effects of three different sperm diluting media, tissue culture medium (TCM-199), sperm culture medium (Sp-TALP) and human tubular fluid (HTF) supplemented with varying concentrations of bovine serum albumin (1, 4 and 6%) or polyvinyl alcohol (0.8%) on the acrosomal integrity, motility and viability of canine spermatozoa. Ejaculates collected from four dogs were diluted in all media and spermatozoa were separated from seminal plasma by the swim-up technique. Sperm progressive motility was assessed using a phase contrast microscope. Viability and acrosomal integrity were evaluated using a dual stain technique (Giemsa-Trypan blue). The results demonstrated that the number of live canine spermatozoa was similar in culture media supplemented or not supplemented with macromolecules. A minimal concentration of albumin (1%) in the three media showed similar effects on vitality, motility and acrosomal integrity, as had higher concentrations (4 and 6%). The percentage of acrosome-intact spermatozoa was markedly higher after HTF (94.1%) than after TCM-199 (70.1%) or Sp-TALP (71.0%) without supplementation. It is concluded that serum bovine albumin, irrespective of the concentration, preserved sperm viability and function, and HTF is the most suitable medium for preserving the acrosome in canine spermatozoa prepared for in vitro manipulation through short incubation.
用于体外受精(IVF)程序的精子培养基是关乎精子活力、运动能力和顶体完整性的重要因素。本研究的目的是调查三种不同的精子稀释培养基,即组织培养基(TCM - 199)、精子培养基(Sp - TALP)和添加不同浓度牛血清白蛋白(1%、4%和6%)或聚乙烯醇(0.8%)的人输卵管液(HTF)对犬精子顶体完整性、运动能力和活力的影响。从四只犬采集的精液在所有培养基中进行稀释,并通过上浮技术将精子与精浆分离。使用相差显微镜评估精子的前向运动能力。使用双重染色技术(吉姆萨 - 台盼蓝)评估活力和顶体完整性。结果表明,添加或未添加大分子物质的培养基中,活犬精子的数量相似。三种培养基中最低浓度的白蛋白(1%)对活力、运动能力和顶体完整性的影响与较高浓度(4%和6%)相似。未添加物质时,HTF处理后顶体完整精子的百分比(94.1%)明显高于TCM - 199(70.1%)或Sp - TALP(71.0%)处理后的百分比。得出的结论是,无论浓度如何,牛血清白蛋白都能保持精子活力和功能,并且HTF是通过短期孵育为体外操作制备的犬精子中保存顶体的最合适培养基。