Department of Production Animal Health, University of Calgary, Calgary, Canada.
Theriogenology. 2011 Jul 15;76(2):252-60. doi: 10.1016/j.theriogenology.2011.02.001. Epub 2011 May 7.
The objective was to evaluate the suitability of an animal protein-free semen extender for cryopreservation of epididymal sperm from the two subspecies of North American bison: plains (Bison bison bison) and wood (Bison bison athabascae) bison. Both cauda epididymides (from six plains and five wood bison) were minced and incubated in Sp-TALPH buffer for approximately 2 h at 37 °C to release actively motile sperm. Sperm suspensions were filtered, centrifuged and the sperm pellet from each bull was divided into two fractions and diluted either in egg yolk containing extender, Triladyl, or in an animal protein-free extender, Andromed, and equilibrated for 20 min at 37 °C. Thereafter, samples were chilled and cryopreserved. Frozen-thawed sperm were evaluated for motility (computer assisted sperm analysis), viability (SYBR 14 and propidium iodide), acrosome integrity (FITC conjugated PSA), cryocapacitation (tyrosine phosphorylation of sperm proteins as a biomarker), and fertilizing ability (in a heterologous IVF system). There was no significant difference for progressive motility, viability, and acrosome integrity between the two extenders for plains bison (36.8 ± 9.0, 60.5 ± 17.4, and 77.3 ± 4.6%; overall mean ± SD) as well as for wood bison (11.7 ± 8.1, 13.7 ± 5.6, and 73.4 ± 4.2%). Levels of tyrosine phosphorylation did not differ for sperm preserved in the two extenders for both subspecies, although an inter-bull variability in the response to tyrosine phosphorylation between extenders was suggested for plains bison. Fertilization percent did not differ significantly between extenders for plains bison (84.16 ± 9.92%, overall mean ± SD) and for wood bison (59.53 ± 19.99%). In conclusion, in the absence of significant difference between extenders in post-thaw sperm characteristics, we inferred that Andromed (animal protein-free) was suitable for cryopreservation of epididymal sperm from North American bison.
本研究旨在评估一种无动物蛋白的精子冷冻液是否适用于北美野牛两个亚种（平原野牛和伍德野牛）的附睾精子冷冻保存。分别从 6 头平原野牛和 5 头伍德野牛的附睾尾部切下小块组织，在 Sp-TALPH 缓冲液中于 37°C 下孵育约 2 小时，以释放具有活力的精子。精子悬液经过过滤和离心后，将每个公牛的精子沉淀分为两部分，分别用含有蛋黄的 Triladyl 或无动物蛋白的 Andromed 稀释液稀释，并在 37°C 下平衡 20 分钟。然后，将样品冷却并冷冻保存。对解冻后的精子进行运动能力（计算机辅助精子分析）、活力（SYBR14 和碘化丙啶）、顶体完整性（FITC 标记的 PSA）、冷冻能力（精子蛋白酪氨酸磷酸化作为生物标志物）和受精能力（在异种 IVF 系统中）进行评估。对于平原野牛，两种稀释液在精子的运动能力、活力和顶体完整性方面没有显著差异（分别为 36.8±9.0%、60.5±17.4%和 77.3±4.6%；总体平均值±标准差），伍德野牛的情况也类似（分别为 11.7±8.1%、13.7±5.6%和 73.4±4.2%）。两种稀释液对两种野牛精子的酪氨酸磷酸化水平均无显著影响，尽管在不同公牛间，两种稀释液对精子的酪氨酸磷酸化反应存在差异。对于平原野牛，两种稀释液的受精率没有显著差异（84.16±9.92%，总体平均值±标准差），对于伍德野牛，两种稀释液的受精率也没有显著差异（59.53±19.99%）。综上所述，在解冻后精子特性方面，两种稀释液没有显著差异，因此我们推断 Andromed（无动物蛋白）适用于北美野牛附睾精子的冷冻保存。
