The aim of this study was to evaluate the applicability of the Cryotech technique for the vitrification of domestic cat (Felis catus) oocytes, as a model for other feline species threatened with extinction. This technique, in which oocytes are stored in a minimal volume of medium, is already widely used in human assisted reproductive technology. In the first part of this study, a viability test (EtBr/FDA) was used to evaluate the toxicity of the vitrification media (solutions). After IVM, oocytes were placed in vitrification and warming solutions according to the manufacturer's procedure, with or without exposure to liquid nitrogen. The solutions and the vitrification procedure each caused a reduction in oocyte viability, with survival rates of 71.4% in oocytes exposed to the Cryotech media (without cooling in liquid nitrogen), and 62% in oocytes that were vitrified. In the second part of the experiment, parthenogenetic activation was used to evaluate the developmental potential of oocytes previously vitrified using the Cryotech method. After warming, the oocytes were activated using a combination of 0.7 µM ionomycin in TCM 199 medium (5 min) followed by 2 mM 6‐DMAP in TCM 199 supplemented with 10% FBS (3 hr), then cultured and evaluated every 24 hr for parthenogenetic cleavage. In the experimental group, 23/50 (46%) cleaved embryos were obtained. Domestic cat oocytes, vitrified by the Cryotech method, are characterized by high survival rates. However, it is necessary to improve the technique to increase the developmental competence of embryos obtained from vitrified oocytes.

中文翻译：

---

Cryotech方法玻璃化后，家猫（Felis catus）卵母细胞的生存能力和发育能力。

这项研究的目的是评估冷冻技术对家猫（Felis catus）玻璃化的适用性。卵母细胞，作为其他濒临灭绝的猫科动物的模型。这种将卵母细胞存储在最小体积的培养基中的技术已经广泛用于人类辅助生殖技术中。在本研究的第一部分中，使用生存力测试（EtBr / FDA）评估了玻璃化介质（溶液）的毒性。IVM后，根据制造商的程序将卵母细胞置于玻璃化和温热的溶液中，有或没有暴露于液氮中。溶液和玻璃化步骤均导致卵母细胞活力降低，暴露于Cryotech培养基（未在液氮中冷却）的卵母细胞的存活率为71.4％，玻璃化的卵母细胞的存活率为62％。在实验的第二部分，单性生殖激活用于评估先前使用Cryotech方法玻璃化的卵母细胞的发育潜力。温热后，使用0.7 µM离子霉素在TCM 199培养基中混合活化卵母细胞（5分钟），然后在补充有10％FBS的TCM 199中使用2 mM 6‐DMAP激活卵母细胞（3小时），然后培养并每24小时评估一次单性生殖分裂。在实验组中，获得了23/50（46％）裂解的胚胎。通过Cryotech方法玻璃化的家猫卵母细胞的特点是存活率高。然而，有必要改进技术来增加从玻璃化卵母细胞获得的胚胎的发育能力。在TCM 199培养基中加入7 µM离子霉素（5分钟），然后在TCM 199中加入2 mM 6–DMAP（补充10％FBS）（3小时），然后每24小时进行培养并评估孤雌分裂。在实验组中，获得了23/50（46％）裂解的胚胎。通过Cryotech方法玻璃化的家猫卵母细胞的特点是存活率高。然而，有必要改进技术来增加从玻璃化卵母细胞获得的胚胎的发育能力。在TCM 199培养基中加入7 µM离子霉素（5分钟），然后在TCM 199中加入2 mM 6–DMAP（补充10％FBS）（3小时），然后每24小时进行培养并评估孤雌分裂。在实验组中，获得了23/50（46％）裂解的胚胎。通过Cryotech方法玻璃化的家猫卵母细胞的特点是存活率高。然而，有必要改进技术来增加从玻璃化卵母细胞获得的胚胎的发育能力。