Moran Sean P, Chi Tim, Prucha Melinda S, Agca Yuksel, Chan Anthony Ws
Division of Neuropharmacology and Neurologic Diseases, Yerkes National Primate Research Center, Atlanta, Georgia, USA.
Division of Neuropharmacology and Neurologic Diseases, Yerkes National Primate Research Center, Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia, USA.
J Am Assoc Lab Anim Sci. 2016;55(5):520-4.
Cryopreservation is an important tool routinely used in preserving sperm for assisted reproductive technologies and for genetic preservation of unique animal models. Here we investigated the viability of fresh and frozen sperm from rhesus macaques on the basis of motility, membrane integrity, and acrosome integrity. Sperm motility was determined by visual evaluation; membrane and acrosome integrity were assessed simultaneously through triple staining with Hoechst 33342, propidium iodide, and fluorescein isothiocyanate-peanut agglutinin. We compared thawed semen that had been cryopreserved by using 2 different media with fresh semen from wildtype (WT) macaques; fresh semen from a model of Huntington disease (HD) with fresh WT semen; and fresh HD with cryopreserved-thawed HD semen. Our new freezing media (TEST EQ) preserved the acrosome better, with less net damage, than did traditional TEST (egg yolk extender containing TES and Tris) media. In addition, the percentage of membrane-damaged cells was similar in fresh HD semen (38.6%±2.9%) and WT semen (35.5%±1.9%). Membrane and acrosomal damage were not different between HD and WT sperm after cryopreservation and subsequent thawing. Furthermore, cryopreservation had similar negative effects on the motility of HD and WT sperm. These data illustrate that semen from a rhesus macaque model of HD is similarly cryotoleratant to that from WT animals.
冷冻保存是一种重要的工具，常用于为辅助生殖技术保存精子以及对独特动物模型进行基因保存。在此，我们基于活力、膜完整性和顶体完整性，研究了恒河猴新鲜精子和冷冻精子的生存能力。精子活力通过视觉评估确定；通过用Hoechst 33342、碘化丙啶和异硫氰酸荧光素 - 花生凝集素进行三重染色，同时评估膜和顶体的完整性。我们将使用两种不同培养基冷冻保存的解冻精液与野生型（WT）猕猴的新鲜精液进行比较；将亨廷顿病（HD）模型的新鲜精液与WT新鲜精液进行比较；以及将新鲜的HD精液与冷冻解冻后的HD精液进行比较。我们的新型冷冻培养基（TEST EQ）比传统的TEST（含有TES和Tris的蛋黄稀释剂）培养基能更好地保存顶体，净损伤更少。此外，新鲜HD精液（38.6%±2.9%）和WT精液（35.5%±1.9%）中膜损伤细胞的百分比相似。冷冻保存并随后解冻后，HD和WT精子之间的膜和顶体损伤没有差异。此外，冷冻保存对HD和WT精子的活力有类似的负面影响。这些数据表明，HD恒河猴模型的精液与WT动物的精液在冷冻耐受性方面相似。
