Centro de Pruebas de Porcino, Área de Investigación Ganadera, Subdirección de Investigación y Tecnología, Instituto Tecnológico Agrario, Consejería de Agricultura y Ganadería, Junta de Castilla y León, Hontalbilla, Segovia, Spain.
Anim Reprod Sci. 2012 Jul;133(1-2):109-16. doi: 10.1016/j.anireprosci.2012.06.010. Epub 2012 Jun 21.
The aim of the present study was to evaluate the cryoprotectant effect of different non-permeating sugars for boar sperm. Pooled semen from three boars was used for the experiments. In the first experiment, the sperm quality of boar sperm cryopreserved with an egg-yolk based extender supplemented with different monosaccharides (glucose, galactose or fructose) was compared to a control cryopreserved in lactose-egg yolk extender. In the second experiment, the effect of five disaccharides (lactose, sucrose, lactulose, trehalose or melibiose) on boar sperm cryosurvival was studied. Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37°C after thawing: percentages of sperm with intact plasma membrane (SIPM), sperm presenting high plasma membrane fluidity (HPMF), sperm with intracellular reactive oxygen substances production (IROSP) and apoptotic sperm (AS). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer-assisted sperm analysis system. Freezing extenders supplemented with each of the monosaccharide presented smaller cryoprotective effect than the control extender supplemented with lactose (P<0.05). However, from the three monosaccharides tested, glucose provided the best sperm quality after freezing-thawing. With respect to the disaccharides studied, samples frozen with the extender supplemented with lactulose exhibited in general the lowest sperm quality, except for the percentage of capacitated sperm, which was highest (P<0.05) in the samples cryopreserved with the trehalose extender. Our results suggest that disaccharides have higher cryoprotective effect than monosaccharides, although the monosaccharide composition of the disaccharides is also important, since the best results were obtained with those disaccharides presenting glucose in their composition.
本研究旨在评估不同非渗透性糖对猪精子的抗冻保护作用。实验使用来自三头公猪的混合精液。在第一个实验中，比较了在含有不同单糖（葡萄糖、半乳糖或果糖）的卵黄基础稀释液中冷冻保存的猪精子与在乳糖-卵黄稀释液中冷冻保存的对照精子的精子质量。在第二个实验中，研究了五种二糖（乳糖、蔗糖、乳果糖、海藻糖或棉子糖）对猪精子冷冻存活的影响。在解冻后 37°C 孵育 30 和 150 分钟时，通过流式细胞术评估了几个精子质量参数：具有完整质膜的精子百分比（SIPM）、具有高质膜流动性的精子百分比（HPMF）、具有细胞内活性氧物质产生的精子百分比（IROSP）和凋亡精子百分比（AS）。此外，在相同的孵育时间内，使用计算机辅助精子分析系统评估了总运动精子（TMS）和渐进性运动精子（PMS）的百分比。与添加乳糖的对照稀释液相比，添加每种单糖的冷冻稀释液的抗冻保护作用较小（P<0.05）。然而，在所测试的三种单糖中，葡萄糖在冷冻和解冻后提供了最佳的精子质量。就研究的二糖而言，与添加海藻糖的稀释液冷冻的样品相比，除了具有最高的顶体反应精子百分比（P<0.05）外，一般来说，添加乳果糖的稀释液冷冻的样品的精子质量最低。我们的结果表明，二糖比单糖具有更高的抗冻保护作用，尽管二糖的单糖组成也很重要，因为在组成中含有葡萄糖的二糖获得了最佳的结果。
