Embryo cryopreservation is an important tool to preserve endangered species. As a cryoprotectant for mouse oocytes, antifreeze protein from Anatolica polita (ApAFP914) has demonstrated utility. In the present study, the effects of controlled slow freezing and vitrification methods on the survival rate of sheep oocytes fertilized in vitro after freezing-thawing were compared. Different ApAFP914 concentrations were added to the vitrification liquid for exploring the effect of antifreeze protein on the warmed embryos. The results showed that the survival and hatching rates of in vitro derived embryos were significantly higher than that of the slow freezing method. Furthermore, among the cryopreserved embryos at different developmental stages, the survival and hatching rates of the expanded blastocyst were significantly higher than those of the blastocysts, early blastocysts and morula. The survival and the hatching rates of the fast-growing embryos were both significantly higher than that of the slow-growing embryos. Additionally, treatment of ApAFP914 (5-30 μg/mL) did not increase the freezing efficiency of the 6-6.5 d embryos. However, addition of 10 μg/mL of ApAFP914 significantly increased the hatching rate of slow-growing embryos. In conclusion, our study suggests that the vitrification is better than the slow freezing method for the conservation of in vitro sheep embryos, and supplementation of ApAFP914 (10 μg/mL) significantly increased the hatching rate of slow-growing embryos after cryopreservation.

中文翻译：

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来自 Anatolia polita 的抗冻蛋白 (ApAFP914) 改善了体外羊胚胎玻璃化的结果

胚胎冷冻保存是保护濒危物种的重要工具。作为小鼠卵母细胞的冷冻保护剂，来自 Anatolica polita 的抗冻蛋白 (ApAFP914) 已被证明是有用的。在本研究中，比较了受控缓慢冷冻和玻璃化方法对冻融后体外受精的羊卵母细胞存活率的影响。将不同浓度的ApAFP914加入玻璃化冷冻液中，研究抗冻蛋白对加热胚胎的影响。结果表明，体外衍生胚胎的存活率和孵化率明显高于慢速冷冻法。此外，在不同发育阶段的冷冻胚胎中，膨胀囊胚的存活率和孵化率明显高于囊胚、早期囊胚和桑椹胚。快速生长胚胎的存活率和孵化率均显着高于缓慢生长胚胎的存活率和孵化率。此外，ApAFP914 (5-30 μg/mL) 的处理不会增加 6-6.5 天胚胎的冷冻效率。然而，添加 10 μg/mL 的 ApAFP914 显着提高了生长缓慢的胚胎的孵化率。总之，我们的研究表明，玻璃化冷冻比慢速冷冻方法更适合体外羊胚胎的保存，并且添加 ApAFP914 (10 μg/mL) 显着提高了冷冻保存后缓慢生长胚胎的孵化率。快速生长胚胎的存活率和孵化率均显着高于缓慢生长胚胎的存活率和孵化率。此外，ApAFP914 (5-30 μg/mL) 的处理不会增加 6-6.5 天胚胎的冷冻效率。然而，添加 10 μg/mL 的 ApAFP914 显着提高了生长缓慢的胚胎的孵化率。总之，我们的研究表明，玻璃化冷冻比慢速冷冻方法更适合体外羊胚胎的保存，并且添加 ApAFP914 (10 μg/mL) 显着提高了冷冻保存后缓慢生长胚胎的孵化率。快速生长胚胎的存活率和孵化率均显着高于缓慢生长胚胎的存活率和孵化率。此外，ApAFP914 (5-30 μg/mL) 的处理不会增加 6-6.5 天胚胎的冷冻效率。然而，添加 10 μg/mL 的 ApAFP914 显着提高了生长缓慢的胚胎的孵化率。总之，我们的研究表明，玻璃化冷冻比慢速冷冻方法更适合体外羊胚胎的保存，并且添加 ApAFP914 (10 μg/mL) 显着提高了冷冻保存后缓慢生长胚胎的孵化率。添加 10 μg/mL ApAFP914 可显着提高生长缓慢的胚胎的孵化率。总之，我们的研究表明，玻璃化冷冻比慢速冷冻方法更适合体外羊胚胎的保存，并且添加 ApAFP914 (10 μg/mL) 显着提高了冷冻保存后缓慢生长胚胎的孵化率。添加 10 μg/mL ApAFP914 可显着提高生长缓慢的胚胎的孵化率。总之，我们的研究表明玻璃化冷冻比慢速冷冻方法对体外绵羊胚胎的保存更好，并且添加 ApAFP914 (10 μg/mL) 显着提高了冷冻保存后缓慢生长胚胎的孵化率。