Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8–15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P > .05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P > .05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P < .05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P < .05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners.

中文翻译：

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使用半吸管或中空纤维装置在简化的封闭系统中对小鼠双细胞和囊胚期胚胎进行玻璃化

双细胞期和囊胚期小鼠胚胎在含有 7.5% 乙二醇 (EG) 和 7.5% 二甲基亚砜 (DMSO) 的培养基中平衡 8-15 分钟。玻璃化在含有 0.5 M 蔗糖和 15% EG + 15% DMSO、17.5% EG + 17.5% DMSO 或 20% EG + 20% DMSO 的培养基中进行 30 秒。然后将它们放在半吸管 (HS) 或中空纤维玻璃化 (HFV) 装置上，并在 0.5 毫升吸管内通过冷却空气进行玻璃化。在双细胞胚胎中，除 20% HS 组外，所有组的存活率均为 100% ( P  > .05)。 除 17.5% 和 20% HFV 组明显低于其他组外，所有玻璃化双细胞组的胚泡发育率与新鲜对照组相似 ( P > .05) < .05)。在囊胚胚胎中，HFV 组分为两个亚组（未塌陷；HFV-NC 和塌陷；HFV-C 囊胚）。15% HFV-NC、17.5% HFV-NC 和 15% HFV-C 组的再扩张率降低 ( P  < .05)，而其余与对照组相似。总之，我们建立了一个简化、可靠和封闭的 HFV 玻璃化半吸管系统，不需要熟练的从业者。