Minimum Volume Vitrification of Immature Feline Oocytes
Minimum Volume Vitrification of Immature Feline Oocytes
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DOI：
10.3791/61523
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发表时间：
2020-06-01
影响因子：
1.2
通讯作者：
Luvoni, Gaia C.
中科院分区：
文献类型：
--
作者：
Colombo, Martina;Luvoni, Gaia C.
在野生动物保护计划中，配子库对于保护有价值个体和稀有物种的遗传资源以及促进生物多样性保护至关重要。在猫科动物中，大多数物种都面临灭绝的威胁，而家养品种被用作提高种质库方案效率的模型。在卵母细胞冷冻保存技术中，玻璃化冷冻在人类和兽医辅助生殖中越来越受欢迎。冷冻玻璃化冷冻最初是为人类卵母细胞和胚胎开发的，现已证明非常适合猫卵母细胞。该方法具有多种优点，例如现场条件的可行性和程序的速度，这在需要处理多个样品时非常有用。然而，效率很大程度上取决于操作员的技能，需要实验室内和实验室间的标准化以及人员培训。该协议描述了在商业支持下对未成熟猫科卵母细胞进行最小体积玻璃化冷冻，从卵母细胞收集到加温的一步步现场友好协议。遵循该协议，尽管在升温后成熟和胚胎发育结果方面仍有改进的空间，但可以预期在升温时保留卵母细胞的完整性和活力（高达 90%）。
In wild animals' conservation programs, gamete banking is crucial to safeguard genetic resources of valuable individuals and rare species and to promote biodiversity preservation. In felids, most species are threatened with extinction, and domestic breeds are used as a model to increase the efficiency of protocols for germplasm banking. Among oocyte cryopreservation techniques, vitrification is more and more popular in human and veterinary assisted reproduction. Cryotop vitrification, which was at first developed for human oocytes and embryos, has demonstrated to be wellsuited for cat oocytes. This method offers several advantages, such as the feasibility in field conditions and the speed of the procedure, which can be helpful when several samples need to be processed. However, the efficiency is strongly dependent on the operator's skills, and intra- and inter-laboratory standardization are needed, as well as personnel training. This protocol describes minimum volume vitrification of immature feline oocytes on a commercial support in a step by step field-friendly protocol, from oocyte collection to warming. Following the protocol, preservation of oocyte integrity and viability at warming (as high as 90%) can be expected, although there is still room for improvement in post-warming maturation and embryonic development outcomes.