鱼类卵巢组织及卵母细胞的冷冻保存研究进展

Recent advances in the studies on cryopreservation of ovarian tissues and oocytes in fish species

  • 摘要: 卵巢组织与卵母细胞的冷冻保存是保护野生鱼类遗传资源和经济鱼类种质资源的重要技术。近年来,各种功能性冷冻保护剂(CPAs)和新型冷冻技术不断涌现,但卵巢细胞的冷冻损伤仍无法完全避免。冷冻保存过程中,样品自身的敏感性、CPAs的选用、脱水降温及解冻复水等过程均会对细胞造成损伤,如冰晶形成、脂质相变及氧化应激等,导致细胞结构和活力受损,进而影响卵巢细胞的存活率和发育潜力。本综述系统介绍了鱼类卵巢组织及卵母细胞冷冻保存技术的发展现状;阐述了冷冻保存过程中细胞所受各类损伤及发生机制,包括机械损伤、氧化应激、代谢异常及遗传物质变异等;并概述了卵巢组织及卵母细胞冷冻保存的优化策略与先进技术。最后,本文就鱼类母系种质库的构建提出展望,旨在进一步揭示卵母细胞冷冻损伤的分子机制以制定针对性优化策略,从而建立高效且标准化的鱼类卵巢组织及卵母细胞冷冻保存技术,助力水产养殖业的可持续发展与濒危水生动物遗传资源的保护。

     

    Abstract: Fish germplasm conservation is crucial for sustainable aquaculture and biodiversity preservation. In contrast to the well-established technique of sperm cryopreservation, the cryopreservation of oocytes remains a major challenge due to their large size, high yolk content, and complex cellular structure. This review systematically synthesizes the current status, persistent challenges, and recent advances in the cryopreservation of fish ovarian tissues and oocytes, with the ultimate aim to provide a theoretical foundation for establishing efficient and standardized cryopreservation protocols. Cryopreservation efficiency for fish ovarian tissues and oocytes is mainly determined by biological characteristics and technical variables. Biologically, the oocyte developmental stage, tissue sample size, and species-specific physiology are crucial. Technically, the composition, concentration, and toxicity of cryoprotectants (CPAs), the choice of cryopreservation protocol, along with the cooling and warming rates, also have significant influence on cellular dehydration, ice crystal formation, survival rate and developmental capacity. The freeze-thaw process and CPAs can induce various cryoinjuries on ovarian cells. These include mechanical damage from ice crystals and membrane lipid phase transition, profound oxidative stress resulting from metabolic disruption and CPA toxicity, as well as a range of cellular and subcellular damage such as disruption of membrane integrity, mitochondrial dysfunction, DNA damage, and chromatin disorganization. To address these challenges, a variety of optimization strategies has been developed. These mainly encompass cellular preconditioning (e.g., membrane lipid modulation, antioxidant application) and targeted mitochondrial protection (e.g., via metabolic inhibition or exogenous supplementation). Concurrently, technological advances focus on ice-crystal control through novel CPAs or ultra-rapid warming, the use of biomaterial-based carriers for enhanced stability, and the adoption of automated platforms like microfluidics to improve standardization and scalability. In conclusion, considerable progress has been made in elucidating cryodamage mechanisms and devising protective strategies for fish ovarian materials. In order to establish a comprehensive fish maternal germplasm biobank, several proposals were advanced, including prioritize studying subcellular cryo-response mechanisms, extending preservation limits, standardizing protocols, and improving in vitro culture systems. Thus, this work would provide the basis for the sustainable development of aquaculture and the conservation of genetic resources in endangered aquatic animals.

     

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