It has been known that different protocols are used for embryo preservation at different stages due to different sensitivity to the physical and physiological stress caused by vitrification. In this study, we developed a common vitrification protocol using carboxlated ε-poly-l-lysine (COOH-PLL), a new cryoprotective agent for the vitrification of mouse embryos at different stages. The IVF-derived Crl:CD1(ICR) x B6D2F1/Crl pronuclear, 2-cell, 4-cell, and 8-cell, morula and blastocyst stage embryos were vitrified with 15% (v/v) ethylene glycol (EG) and 10% (w/v) COOH-PLL (E15P15) or 15% (v/v) EG and 15% (v/v) dimethyl sulfoxide (E15D15) using the minimal volume cooling method. The survival of vitrified embryos from pronuclear to blastocyst stages was equivalent between E15P15 and E15D15 groups. However, the rate of development to blastocysts was significantly lower in E15P15 than E15D15. The rates of survival and development to blastocysts were dramatically improved by a slight modification of EG and COOH-PLL concentrations (E20P10). After transferring 17 (E20P10) and 15 (E15D15) vitrified/warmed blastocysts, 8 and 7 pups were obtained (47.1% and 46.7%, respectively). Taken together, these results indicate that our vitrification protocol is appropriate for the vitrification of mouse embryos at different stages.

中文翻译：

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羧化 ε-聚-L-赖氨酸，一种冷冻保护剂，是乙二醇的有效搭档，用于胚胎植入前各个阶段的玻璃化

众所周知，由于对玻璃化造成的物理和生理压力的敏感性不同，不同阶段的胚胎保存采用不同的方案。在这项研究中，我们开发了一种使用羧化 ε-聚-l-赖氨酸 (COOH-PLL) 的通用玻璃化方案，这是一种用于不同阶段小鼠胚胎玻璃化的新型冷冻保护剂。IVF 衍生的 Crl:CD1(ICR) x B6D2F1/Crl 前核、2 细胞、4 细胞和 8 细胞、桑椹胚和囊胚期胚胎用 15% (v/v) 乙二醇 (EG) 和10% (w/v) COOH-PLL (E15P15) 或 15% (v/v) EG 和 15% (v/v) 二甲基亚砜 (E15D15) 使用最小体积冷却方法。E15P15 和 E15D15 组之间从原核到胚泡阶段的玻璃化胚胎的存活率相等。然而，E15P15 的胚泡发育率明显低于 E15D15。通过略微修改 EG 和 COOH-PLL 浓度 (E20P10)，显着提高了胚泡的存活率和发育率。转移 17 个 (E20P10) 和 15 个 (E15D15) 玻璃化/加热的囊胚后，获得了 8 只和 7 只幼崽（分别为 47.1% 和 46.7%）。总之，这些结果表明我们的玻璃化方案适用于不同阶段的小鼠胚胎玻璃化。