Background Although sperm cryopreservation has been widely used in human reproductive medicine as an integral infertility management in infertility clinics and for banking sperm in sperm banks, the freezing/thawing protocols are not optimal. The freezing and thawing processes result in changes at both structural and molecular levels, some even detrimental, in human sperm when compared with fresh sperm. The change of sperm proteins after cryopreservation may play negative roles for fertilization and early embryo development. Conventionally, cryostraws (CS) and cryovials (CV) are the most widely used cryopreservation carriers (CPCs) for human sperm cryopreservation accompanied with the use of egg yolk free commercial media. However, the influence of cryopreservation on the proteomic profile of human sperm preserved with the two CPCs is unknown. Therefore the purpose of the present study was to compare the frozen-thawed motility, investigate the proteomic profile of human sperm cryopreserved with the two types of CPCs, and identify the susceptible proteins that play key roles for sperm function and fertility. Methods The present study compared the cryosurvival of human sperm frozen with the two different CPCs and identified the sperm proteomic changes by using the isobaric tags for relative and absolute quantification labeling technique coupled with 2D LC-MS/MS analysis after freezing and thawing. Results Our results indicated that sperm cryopreserved with CV showed higher values for percentage of motile sperm and forward activity rate than those with CS. Compared to fresh sperm, 434 and 432 proteins were differentially identified in human sperm cryopreserved with CS and CV, respectively. Conclusion The proteomic profiles of human sperm are greatly affected by cryopreservation with either type of CPC. GO analysis revealed that most of the differentially identified sperm proteins enriched in the extracellular membrane-bounded organelles, cytoplasm and cytosol. In addition, 106 susceptible proteins having known identities related to sperm functions were identified. In general, cryovial seems to be the preferred CPC for human sperm cryopreservation based on the post-thaw motility parameters and the effect on sperm proteomic profiles. These results are beneficial for the insight into the understanding of the cryoinjury mechanism of sperm and the development of human sperm cryopreservation strategies.

中文翻译：

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用冷冻管和冷冻管冷冻保存的人类精子的差异运动参数和蛋白质组学特征的鉴定。

背景 虽然精子冷冻保存已广泛用于人类生殖医学，作为不孕症诊所的整体不孕症管理以及将精子储存在精子库中，但冷冻/解冻方案并不是最佳的。与新鲜精子相比，冷冻和解冻过程会导致人类精子在结构和分子水平上发生变化，有些甚至是有害的。冷冻保存后精子蛋白的变化可能对受精和早期胚胎发育产生不利影响。传统上，冷冻管 (CS) 和冷冻管 (CV) 是用于人类精子冷冻保存的最广泛使用的冷冻保存载体 (CPC)，同时使用无蛋黄的商业介质。然而，冷冻保存对两种 CPC 保存的人类精子蛋白质组学特征的影响尚不清楚。因此，本研究的目的是比较冻融后的运动能力，研究用两种 CPCs 冷冻保存的人类精子的蛋白质组学特征，并确定对精子功能和生育能力起关键作用的易感蛋白质。方法本研究比较了两种不同CPCs冷冻人类精子的冷冻存活率，并通过使用等压标签进行相对和绝对定量标记技术结合2D LC-MS/MS分析冷冻和解冻后的精子蛋白质组变化。结果 我们的研究结果表明，CV 冷冻精子的活动精子百分比和前向活动率高于 CS。与新鲜精子相比，在用 CS 和 CV 冷冻保存的人类精子中差异鉴定了 434 和 432 种蛋白质，分别。结论 人类精子的蛋白质组学特征受任何一种 CPC 冷冻保存的影响很大。GO分析显示，大多数差异鉴定的精子蛋白富含细胞外膜结合的细胞器、细胞质和胞质溶胶。此外，还鉴定了 106 种具有与精子功能相关的已知特性的易感蛋白质。一般来说，根据解冻后的运动参数和对精子蛋白质组学特征的影响，冷冻管似乎是人类精子冷冻保存的首选 CPC。这些结果有利于深入了解精子的冷冻损伤机制和人类精子冷冻保存策略的发展。GO分析显示，大多数差异鉴定的精子蛋白富含细胞外膜结合的细胞器、细胞质和胞质溶胶。此外，还鉴定了 106 种具有与精子功能相关的已知特性的易感蛋白质。一般来说，根据解冻后的运动参数和对精子蛋白质组学特征的影响，冷冻管似乎是人类精子冷冻保存的首选 CPC。这些结果有利于深入了解精子的冷冻损伤机制和人类精子冷冻保存策略的发展。GO分析显示，大多数差异鉴定的精子蛋白富含细胞外膜结合的细胞器、细胞质和胞质溶胶。此外，还鉴定了 106 种具有与精子功能相关的已知特性的易感蛋白质。一般来说，根据解冻后的运动参数和对精子蛋白质组学特征的影响，冷冻管似乎是人类精子冷冻保存的首选 CPC。这些结果有利于深入了解精子的冷冻损伤机制和人类精子冷冻保存策略的发展。鉴定了 106 种具有与精子功能相关的已知特性的易感蛋白质。一般来说，根据解冻后的运动参数和对精子蛋白质组学特征的影响，冷冻管似乎是人类精子冷冻保存的首选 CPC。这些结果有利于深入了解精子的冷冻损伤机制和人类精子冷冻保存策略的发展。鉴定了 106 种具有与精子功能相关的已知特性的易感蛋白质。一般来说，根据解冻后的运动参数和对精子蛋白质组学特征的影响，冷冻管似乎是人类精子冷冻保存的首选 CPC。这些结果有利于深入了解精子的冷冻损伤机制和人类精子冷冻保存策略的发展。