Dipartimento di Scienze Veterinarie per la Salute, la Produzione Animale e la Sicurezza Alimentare "Carlo Cantoni", Università degli Studi di Milano, Via Celoria 10, 20133 Milan, Italy.
Department of Reproduction Biology, Leibniz-Institute for Zoo and Wildlife Research, Alfred-Kowalke-Str. 17, D-10315, Berlin, Germany.
Cryobiology. 2021 Feb;98:181-186. doi: 10.1016/j.cryobiol.2020.11.003. Epub 2020 Nov 7.
In feline species, cooled transport of ovaries can be employed without detrimental effects to retrieve immature oocytes intended for in vitro embryo production purposes. Indeed, this is the most common way to collect gametes from gonads of wild, valuable animals after they die or are castrated far from specialized laboratories. However, fresh retrieved gametes are generally used, and their cryosensitivity is not known. This study employed ovariectomy-derived domestic cat gonads as a model for wild felids, and aimed to compare the yield and developmental competence of Cryotop-vitrified oocytes (VOs) collected and cryopreserved right after ovary excision (In loco-VOs) or after 24 h cooled transport of ovaries (Shipped-VOs). The number of collected oocytes was higher in In loco-VOs than in Shipped-VOs (mean ± SD: 8 ± 3.36 vs 5.6 ± 3.1, p = 0.05). In vitro embryo production resulted in similar maturation (35% for both vitrified groups, p = 1) and fertilization rates (In loco-VOs: 29.1%; Shipped-VOs: 22.2%; p = 0.295), but showed a difference in cleavage (In loco-VOs: 25.6%; Shipped-VOs: 14.5%; p = 0.0495). No differences were found in further embryo development. Taken together, results suggested that delayed oocyte vitrification after cooled transport of organs was feasible and allowed embryo development. However, the number of collected oocytes and the cleavage rate of matured oocytes were higher when oocyte vitrification was performed without delay after ovary excision, and this should be considered in gamete conservation programs for endangered felids.
在猫科动物中,可以采用冷却运输卵巢的方法,不会对卵巢造成损害,从而获取用于体外胚胎生产目的的未成熟卵母细胞。事实上,这是从死亡或远离专门实验室的野生动物性腺中收集配子的最常见方法。然而,通常使用新鲜采集的配子,并且其对冷冻的敏感性尚不清楚。本研究以卵巢切除术衍生的家猫性腺作为野生猫科动物的模型,旨在比较直接在卵巢切除后(loco-VOs)或卵巢冷却运输 24 小时后(Shipped-VOs)采集和冷冻保存的 Cryotop 玻璃化卵母细胞(VOs)的产量和发育能力。In loco-VOs 中收集的卵母细胞数量高于 Shipped-VOs(平均值 ± 标准差:8 ± 3.36 对 5.6 ± 3.1,p = 0.05)。体外胚胎生产导致相似的成熟率(两个玻璃化组均为 35%,p = 1)和受精率(In loco-VOs:29.1%;Shipped-VOs:22.2%;p = 0.295),但卵裂率存在差异(In loco-VOs:25.6%;Shipped-VOs:14.5%;p = 0.0495)。进一步的胚胎发育没有差异。综上所述,结果表明,在器官冷却运输后延迟卵母细胞玻璃化是可行的,并允许胚胎发育。然而,当卵母细胞在卵巢切除后不延迟进行玻璃化时,采集的卵母细胞数量和成熟卵母细胞的卵裂率更高,这在濒危猫科动物的配子保存计划中应该考虑到。