普通狨猴精子处理与冷冻保存流程优化:微流控提升活力并减少DNA损伤
Analysis of sperm processing and cryopreservation procedures in the common marmoset ( Callithrix jacchus) to reduce fragmentation and maximize total and progressive motility
摘要
该预印本研究以普通狨猴为对象,比较四种孵育前处理及上游法、蔗糖梯度与微流控三种精子处理方式,并用CASA评估精子参数。结果显示38°C TYH培养基孵育30分钟可减少精子聚集,后续微流控处理的总活力与前向运动比例优于上游法和梯度法。在五种冷冻保存方案中,优化方案解冻后稳定达到至少20%前向运动,且未造成DNA损伤。研究为狨猴IVF与基因编辑相关精子处理和冻存提供了标准化参考。
原文摘要
The marmoset has emaserged as an ideal species for studying complex neurodevelopmental diseases, and IVF is required for gene-editing success. Reliable sperm processing is paramount to several IVF techniques, including traditional insemination. If not used immediately, thermal stress from cryopreservation diminishes sperm quality, requiring improved processing methods. Swim-up and gradient methods may limit adequate sperm quality, and the lack of a standardized freezing protocol often results in impaired post-thaw motility. Thus, reliable standards for sperm processing and cryopreservation in the marmoset are needed to support vital IVF processes. Collected semen underwent one of four pre-processing incubation steps before processing by swim-up, sucrose gradient, or a microfluidics assay. Sperm parameters were assessed using a Hamilton Thorne Computer Assisted Sperm Analysis (CASA) system. To evaluate cryopreservation, microfluidics-processed sperm were subjected to five protocols using different cryoprotectants and cooling methods. After thawing, sperm samples were analyzed with the CASA system and a DNA fragmentation assay. Incubation for 30 minutes at 38°C in TYH media decreased sperm aggregation, and subsequent microfluidics resulted in a higher percentage of motile and progressively motile sperm than swim-up or gradient methods. Our optimized cryopreservation protocol consistently achieved at least 20% progressive motility, a key benchmark for usable marmoset sperm, while previously published or commercial methods were less consistent. The cryopreservation protocol outperformed other published methods, having the least significant change in post-thaw motility without causing DNA damage. These findings establish standardized methods for reliably procuring and storing sperm samples for IVF or husbandry in the common marmoset.
推荐理由:对狨猴基因编辑与IVF实验室的精子处理、冻存标准化有直接实用价值,微流控与冻存方案比较设计较清晰;但属预印本,样本量与统计细节可见信息有限,对人类ART的直接外推价值有限。
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所属事件:普通狨猴精子处理与冷冻保存流程优化:微流控提升活力并减少DNA损伤 热度 3.6 · 3 个来源
收录来源(3)
- 11-08 11:43PubMed · 辅助生殖临床
- 11-08 11:43PubMed · 配子与性腺
- 11-08 11:44Europe PMC · 预印本(领域内)