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DOI: 10.3791/68176-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
在这里,我们提出了用于玻璃化小鼠和绵羊植入前胚胎的 Spatula Montevideo 方案,包括自制刮刀和利用自制解决方案。胚胎在两种浓度递增的冷冻保护剂溶液中快速平衡,加载到抹刀上,浸入液氮中,最后用 0.5 mL 吸管密封。
生成转基因小鼠模型,然后冷冻保存品系作为备份。为此,我们需要一种简单、低成本、高效且可重复的方法来玻璃化胚胎。CRISPR-Cas9的显微注射或电穿孔可以在玻璃化、预热小鼠和牲畜受精卵中进行,从而实现更灵活的时间表并减少动物的使用。
胚胎玻璃化使用超快速冷却来防止冰晶形成,保持细胞完整性。该技术推进了冷冻生物学研究,使胚胎活力、遗传稳定性和长期储存结果的研究成为可能。抹刀 MVD 和溶液都是自制的。抹刀用 0.5 毫升吸管密封,在液氮杜瓦瓶中占用的空间最小。
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