Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Cryobiology. 2020 Feb 1;92:208-214. doi: 10.1016/j.cryobiol.2020.01.013. Epub 2020 Jan 28.
The freeze-thaw procedure causes irreversible structural and functional changes in human spermatozoa. In order to decrease the detrimental effects of cryopreservation and improve the quality of post-thawed spermatozoa, the constituents of the freezing solution attracted considerable attention. In this study, for the first time, we evaluated the efficacy of knockout serum replacement (KSR) as a substitute for human serum albumin (HSA) for cryopreservation of human spermatozoa. Twenty semen samples were collected from normozoospermic men and divided them into five equal groups. One of the aliquots was diluted with glycerol-based medium as a control group (CON). The other four aliquots were diluted with the sucrose solution containing 5% HSA (H5), 10% HSA (H10), 5% KSR (K5), and 10% KSR (K10). The diluted samples were frozen and preserved in liquid nitrogen. Post thawed sperm parameters including motion characteristics, viability, membrane integrity, mitochondrial activity, acrosome integrity and DNA intactness in all of the sucrose-based groups were comparable with glycerol-based medium. The replacement of HSA by 10% KSR in the freezing medium resulted in significantly higher post-thawed viability, acrosome integrity and DNA intactness compared with other sucrose-based groups. In conclusion, the addition of 10% KSR to the sucrose-based freezing solution improves the quality of post-thawed human spermatozoa and may have potential to develop chemically defined freezing medium.
冻融过程会导致人类精子不可逆的结构和功能变化。为了降低冷冻保存的有害影响，提高解冻后精子的质量，冷冻液的成分引起了人们的极大关注。在这项研究中，我们首次评估了无血清替代物（KSR）作为人血清白蛋白（HSA）替代物用于人类精子冷冻保存的效果。从正常精子的 20 份精液样本中随机抽取，分为五组。其中一份用甘油基培养基稀释作为对照组（CON）。另外四份用含有 5% HSA（H5）、10% HSA（H10）、5% KSR（K5）和 10% KSR（K10）的蔗糖溶液稀释。稀释后的样本在液氮中冷冻保存。所有基于蔗糖的组的解冻后精子参数，包括运动特性、活力、膜完整性、线粒体活性、顶体完整性和 DNA 完整性，与甘油基培养基相当。与其他基于蔗糖的组相比，冷冻液中用 10% KSR 替代 HSA 可显著提高解冻后精子的活力、顶体完整性和 DNA 完整性。总之，在蔗糖基冷冻液中添加 10% KSR 可提高解冻后人类精子的质量，并可能有潜力开发化学定义的冷冻液。
