Gutiérrez-Pérez Oscar, Juárez-Mosqueda María de Lourdes, Carvajal Salvador Uribe, Ortega María Elena Trujillo
Department of Pig Production, FMVZ, Universidad Nacional Autónoma de México, México DF, CP, Mexico.
Cryobiology. 2009 Jun;58(3):287-92. doi: 10.1016/j.cryobiol.2009.02.003. Epub 2009 Feb 20.
The use of glycerol for boar semen cryopreservation results in low fertility, possibly due to toxicity. This has led to recommend the use of solutions with less than 4% glycerol. Trehalose is a disaccharide known to stabilize proteins and biologic membranes during processes such as cryopreservation. Thus, it was decided to evaluate the cryoprotective effect of glycerol/trehalose mixtures. Effects on motility (M), viability (Vb) and acrosomal integrity (nA) were evaluated. Sperm samples were frozen in three different extenders: G4 contained 4% glycerol; T1 contained 1% glycerol plus 250 mM trehalose and T0.5 was constituted by 0.5% glycerol plus 250 mM trehalose. All extenders yielded similar post-freezing/thawing motility rates. Viability was diminished in T0.5 as compared to the others. In regard to acrosome integrity, it was twice as high (P<0.05) in the trehalose enriched media as in G4, the glycerol-only extender. Thus, T1 twice as many spermatozoa were alive, motile and intact, than in either T0.5 or G4, i.e. during freeze/thawing the use of T1 resulted in twice as many fertile cells as when using the other extenders. During our study, we noted that there were wide individual variations both in sperm viability and in motility.
使用甘油对公猪精液进行冷冻保存会导致生育力低下，这可能是由于毒性所致。因此，有人建议使用甘油含量低于4%的溶液。海藻糖是一种二糖，已知在冷冻保存等过程中能稳定蛋白质和生物膜。因此，决定评估甘油/海藻糖混合物的冷冻保护作用。评估了对精子活力（M）、存活率（Vb）和顶体完整性（nA）的影响。精子样本在三种不同的稀释液中冷冻：G4含有4%的甘油；T1含有1%的甘油加250 mM海藻糖，T0.5由0.5%的甘油加250 mM海藻糖组成。所有稀释液在冷冻/解冻后的活力率相似。与其他组相比，T0.5组的存活率有所降低。关于顶体完整性，富含海藻糖的培养基中的顶体完整性是仅含甘油的G4培养基中的两倍（P<0.05）。因此，T1组存活、有活力且完整的精子数量是T0.5组或G4组的两倍，即在冷冻/解冻过程中，使用T1组产生的可育细胞数量是使用其他稀释液时的两倍。在我们的研究过程中，我们注意到精子的存活率和活力都存在很大的个体差异。
