Mahiddine Feriel Yasmine, Kim Jin Wook, Qamar Ahmad Yar, Ra Jeong Chan, Kim Soo Hyun, Jung Eun Joong, Kim Min Jung
Department of Theriogenology and Biotechnologies, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea.
Laboratory of Theriogenology, College of Veterinary Medicine, Chungnam National University, Daejeon 34134, Korea.
Animals (Basel). 2020 Oct 16;10(10):1899. doi: 10.3390/ani10101899.
This study investigated the effects of conditioned medium (CM) from canine amniotic membrane-derived MSCs (cAMSCs) on dog sperm cryopreservation. For this purpose, flow cytometry analysis was performed to characterize cAMSCs. The CM prepared from cAMSCs was subjected to proteomic analysis for the identification of proteins present in the medium. Sperm samples were treated with freezing medium supplemented with 0%, 5%, 10%, and 15% of the CM, and kinetic parameters were evaluated after 4-6 h of chilling at 4 °C to select the best concentration before proceeding to cryopreservation. Quality-related parameters of frozen-thawed sperm were investigated, including motility; kinetic parameters; viability; integrity of the plasma membrane, chromatin, and acrosome; and mitochondrial activity. The results showed that 10% of the CM significantly enhanced motility, viability, mitochondrial activity, and membrane integrity ( < 0.05); however, the analysis of chromatin and acrosome integrity showed no significant differences between the treatment and control groups. Therefore, we concluded that the addition of 10% CM derived from cAMSC in the freezing medium protected dog sperm during the cryopreservation process.
本研究调查了犬羊膜来源间充质干细胞（cAMSCs）的条件培养基（CM）对犬精子冷冻保存的影响。为此，进行了流式细胞术分析以表征cAMSCs。对从cAMSCs制备的CM进行蛋白质组学分析，以鉴定培养基中存在的蛋白质。精子样本用补充有0%、5%、10%和15% CM的冷冻培养基处理，并在4℃冷藏4-6小时后评估动力学参数，以选择最佳浓度，然后再进行冷冻保存。研究了冻融精子的质量相关参数，包括活力；动力学参数；存活率；质膜、染色质和顶体的完整性；以及线粒体活性。结果表明，10%的CM显著提高了活力、存活率、线粒体活性和膜完整性（<0.05）；然而，染色质和顶体完整性分析显示，处理组和对照组之间没有显著差异。因此，我们得出结论，在冷冻培养基中添加10%源自cAMSC的CM可在冷冻保存过程中保护犬精子。
