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Development of Cryopreservation Techniques for Gorgonian (Junceella juncea) Oocytes through Vitrification

机译:玻璃化冷冻高粱(Junceella juncea)卵母细胞冷冻保存技术的发展

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摘要

Gorgonian corals are slowly declining due to human interaction and environmental impacts. Cryopreservation of gorgonian corals is an ex-situ method of conservation, ensuring future reproduction. The present study assessed the vitrification properties of cryoprotectant (CPT) mixtures using the cryotop, cryoloop and open pulled straw (OPS) cryopereservation methods prior to experimentation on gorgonian (Junceella juncea) oocytes. Investigations of the equilibration and vitrification solutions’ (ES and VS) effect on oocytes throughout different incubation periods were conducted. The cryotop method was found to be the most successful in ensuring vitrification. The most favourable VS was composed of propylene glycol (PG), ethylene glycol (EG) and methanol with concentrations of 3.5M, 1.5M and 2M respectively. Experiments were performed using the cryotop method to cryopreserve Junceella juncea oocytes using VS2, the solution had the least impact on oocytes at 5°C rather than at 26°C. The success of the vitrification procedures was determined by adenosine triphosphate (ATP) levels in cooled-thaw oocytes and the highest viability obtained from the present study was 76.6 ± 6.2%. This study provides information regarding gorgonian corals’ tolerance and viability throughout vitrification to further advance the vitrification protocol on whip corals.
机译:由于人类之间的相互作用和环境影响,古高阶珊瑚正在缓慢下降。冷冻珊瑚是一种非原生境的保护方法,可确保将来的繁殖。本研究评估了冷冻保护剂(CPT)混合物的玻璃化特性,然后在对高粱(Junceella juncea)卵母细胞进行实验之前,先使用了cryotop,cryoloop和开放式吸管(OPS)冷冻保存方法。研究了平衡和玻璃化溶液(ES和VS)在整个潜伏期对卵母细胞的影响。发现cryotop方法是确保玻璃化的最成功方法。最有利的VS由丙二醇(PG),乙二醇(EG)和甲醇组成,其浓度分别为3.5M,1.5M和2M。使用cryotop方法进行实验,使用VS2冷冻junceella juncea卵母细胞,该溶液在5°C而不是26°C对卵母细胞的影响最小。玻璃化程序的成功由冷却融化的卵母细胞中的三磷酸腺苷(ATP)水平决定,从本研究获得的最高生存力为76.6±6.2%。这项研究提供了有关整个玻璃化过程中珊瑚的耐受性和生存能力的信息,以进一步推进鞭状珊瑚的玻璃化协议。

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