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 (p < 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.

中文翻译：

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犬羊膜来源间充质干细胞的条件培养基改善狗精子解冻后质量相关参数

这项研究调查了来自犬羊膜来源的MSC（cAMSC）的条件培养基（CM）对狗精子冷冻保存的影响。为此目的，进行了流式细胞术分析以表征cAMSC。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。精子样品用添加了0％，5％，10％和15％CM的冷冻培养基处理，并在4°C冷冻4–6 h后评估动力学参数，以选择最佳浓度，然后再进行冷冻保存。研究了与冻融精子质量相关的参数，包括运动性；动力学参数 可行性; 质膜，染色质和顶体的完整性；和线粒体活性。p <0.05）；然而，染色质和顶体完整性的分析显示，治疗组和对照组之间没有显着差异。因此，我们得出结论，在冷冻保存过程中，在冷冻培养基中添加了cAMSC衍生的10％CM保护了狗的精子。