Laboratory of Reproductive Medicine and Molecular Endocrinology, Center for Translational Medicine (CEMT-BIOREN), Universidad de La Frontera, Temuco, Chile; Escuela de Medicina Veterinaria, Facultad de Recursos Naturales y Medicina Veterinaria, Universidad Santo Tomas, Chile.
Laboratory of Reproductive Medicine and Molecular Endocrinology, Center for Translational Medicine (CEMT-BIOREN), Universidad de La Frontera, Temuco, Chile; Department of Preclinical Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco, Chile.
Cryobiology. 2020 Dec;97:5-11. doi: 10.1016/j.cryobiol.2020.10.015. Epub 2020 Oct 27.
Saccharides have bioprotective properties, with a high capacity to preserve biological proteins and membranes during sperm cryopreservation. The aim of this study was to evaluate how replacing the lactose of cryopreservation media by sucrose (SUC) or trehalose (TRE) at concentrations of 0.2 M (SUC-1 and TRE-1) and 0.25 M (SUC-2 and TRE-2) affects frozen/thawed pig spermatozoa. The media used were composed of medium A (saccharide/egg yolk) and B (saccharide/egg yolk/glycerol), their osmolality being determined prior to freezing. Cell viability, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), lipid peroxidation, thiol group oxidation, total reactive oxygen species (ROS), peroxynitrite and superoxide anion (O) were determined through flow cytometry; total motility (TM), progressive motility (PM) and kinetic parameters motility were determined immediately after thawing (T0) and again 30 (T30) and 60 (T60) minutes later. The SUC-2 and TRE-2 groups maintained viability significantly and presented fewer lipid membrane disorders, respectively, both with a significant increase in MMP. The production of O and peroxynitrite was lower in the TRE-2 groups compared to the control (P < 0.05). Total motility at T0 was greater in the TRE-2 group (P < 0.05). Sperm kinetics was not affected by the treatment. The use of saccharides SUC and TRE at a concentration of 0.25 M improves sperm quality, so that both non-penetrating cryoprotectants can be utilized in pig sperm freezing media.
糖具有生物保护特性，在精子冷冻保存过程中，具有高能力来保护生物蛋白质和膜。本研究的目的是评估在 0.2M（SUC-1 和 TRE-1）和 0.25M（SUC-2 和 TRE-2）浓度下用蔗糖（SUC）或海藻糖（TRE）替代冷冻保存介质中的乳糖对冷冻/解冻猪精子的影响。使用的介质由 A 介质（糖/蛋黄）和 B 介质（糖/蛋黄/甘油）组成，在冷冻前测定其渗透压。通过流式细胞术测定细胞活力、膜脂质紊乱、顶体完整性、线粒体膜电位（MMP）、脂质过氧化、硫醇基团氧化、总活性氧（ROS）、过氧亚硝酸盐和超氧阴离子（O）；总运动性（TM）、进行性运动性（PM）和运动性动力学参数在解冻后立即（T0）以及 30 分钟（T30）和 60 分钟（T60）后测定。SUC-2 和 TRE-2 组的活力保持显著，脂质膜紊乱分别显著减少，MMP 均显著增加。与对照组相比，TRE-2 组的 O 和过氧亚硝酸盐的产生较低（P<0.05）。TRE-2 组的 T0 总运动性较大（P<0.05）。处理对精子动力学没有影响。使用浓度为 0.25M 的 SUC 和 TRE 可改善精子质量，因此这两种非穿透性冷冻保护剂都可用于猪精子冷冻保存介质。
