Rota A, Penzo N, Vincenti L, Mantovani R
Istituto di Patologia e Igiene Veterinaria, Faculty of Veterinary Medicine, University of Padua, Italy.
Theriogenology. 2000 Apr 15;53(7):1415-20. doi: 10.1016/s0093-691x(00)00284-3.
Semen from 5 Piedmontese bulls was subjected to the hypoosmotic swelling (HOS) test in order to determine if the results could be correlated to the fertilizing capacity in vitro. Semen was routinely prepared for in vitro fertilization (IVF), with aliquots being sampled after thawing, after separation on a Percoll gradient and after capacitation in a medium containing heparin. The aliquots were added to a fructose-sodium citrate hypoosmotic solution (100 mOsm) and incubated at 37 degrees C for 5 min. At least 200 spermatozoa were observed at x 400 and classified according to the presence or the absence of a swollen tail. After capacitation, spermatozoa were used to fertilize in vitro-matured bovine oocytes (1.5 x10(6) cells/mL); IVF was performed in Fert-TALP medium supplemented with 6 mg/mL BSA and 10 microL/mL heparin in a humidified atmosphere with 5% CO2 and 5% oxygen. Presumptive zygotes were cultured in SOF medium supplemented with 8 mg/mL BSA and amino acids. There were no significant differences in the in vitro fertility of the bulls, but a significant difference was found between bulls in the response to the HOS test. The 3 assays were significantly correlated, while no significant correlation was observed between the percentage of swollen spermatozoa and in vitro fertility. The HOS test does not appear to be sufficiently sensitive to discriminate between semen samples of intermediate fertility like those used in this preliminary research.
对5头皮埃蒙特公牛的精液进行低渗肿胀（HOS）试验，以确定结果是否与体外受精能力相关。精液按常规方法制备用于体外受精（IVF），在解冻后、在Percoll梯度上分离后以及在含肝素的培养基中获能后进行等分取样。将等分试样加入果糖 - 柠檬酸钠低渗溶液（100 mOsm）中，并在37℃下孵育5分钟。在400倍放大倍数下观察至少200个精子，并根据有无肿胀的尾巴进行分类。获能后，精子用于使体外成熟的牛卵母细胞受精（1.5×10⁶个细胞/mL）；IVF在补充有6 mg/mL BSA和10 μL/mL肝素的Fert-TALP培养基中，在含有5% CO₂和5%氧气的湿润气氛中进行。推定的受精卵在补充有8 mg/mL BSA和氨基酸的SOF培养基中培养。公牛的体外受精能力没有显著差异，但在公牛对HOS试验的反应方面发现了显著差异。这三种测定方法显著相关，而肿胀精子的百分比与体外受精能力之间未观察到显著相关性。HOS试验似乎不够灵敏，无法区分像本初步研究所用的中等受精能力的精液样本。
