High osmolality vitrification: a new method for the simple and temperature-permissive cryopreservation of mouse embryos.
High osmolality vitrification: a new method for the simple and temperature-permissive cryopreservation of mouse embryos.
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DOI：
10.1371/journal.pone.0049316
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ogura A
中科院分区：
文献类型：
--
作者：
Mochida K;Hasegawa A;Li MW;Fray MD;Kito S;Vallelunga JM;Lloyd KC;Yoshiki A;Obata Y;Ogura A
冷冻保存胚胎的程序差异很大，每种程序在技术可行性，胚胎存活率，温度允许性和物种或品系依赖性适用性方面都有其特定的优点和缺点。在这里，我们报告了高渗透压玻璃化（HOV）的方法，在这些方面是有利的。玻璃化冷冻保存通常非常简单，但与缓慢冷冻不同，胚胎应保持在过冷温度（低于-130 ° C）以避免冷冻损伤。我们通过使用含有42.5%（v/v）乙二醇、17.3%（w/v）Ficoll和1.0M蔗糖的HOV溶液克服了这个问题。该溶液比其他冷冻保存溶液更粘稠，但通过在玻璃化前采用较低粘度的平衡溶液来确保胚胎的容易处理。大多数（>80%）在这种溶液中冷冻保存的胚胎在-80 ° C下存活至少30天。正常小鼠即使在常规干冰包装中洲际运输2-3天后也能恢复，这表明不需要特殊容器，如装有液氮蒸汽的干燥运输箱。HOV溶液也可以用于在液氮中长期储存，与其他常规冷冻保护剂一样。最后，我们证实这种新的玻璃化冷冻方法可以成功地应用于我们迄今为止测试的所有六种小鼠的胚胎。因此，我们的HOV方法为动物设施和生物医学实验室中小鼠胚胎的常规冷冻保存提供了一种有效和可靠的策略，并且便于廉价运输。
Procedures for cryopreserving embryos vary considerably, each having its specific advantages and disadvantages in terms of technical feasibility, embryo survival yield, temperature permissibility and species- or strain-dependent applicability. Here we report a high osmolality vitrification (HOV) method that is advantageous in these respects. Cryopreservation by vitrification is generally very simple, but, unlike slow freezing, embryos should be kept at a supercooling temperature (below –130°C) to avoid cryodamage. We overcame this problem by using an HOV solution containing 42.5% (v/v) ethylene glycol, 17.3% (w/v) Ficoll and 1.0 M sucrose. This solution is more viscous than other cryopreservation solutions, but easy handling of embryos was assured by employing a less viscous equilibration solution before vitrification. Most (>80%) embryos cryopreserved in this solution survived at –80°C for at least 30 days. Normal mice were recovered even after intercontinental transportation in a conventional dry-ice package for 2–3 days, indicating that special containers such as dry shippers with liquid nitrogen vapor are unnecessary. The HOV solution could also be employed for long-term storage in liquid nitrogen, as with other conventional cryoprotectants. Finally, we confirmed that this new vitrification method could be applied successfully to embryos of all six strains of mice we have tested so far. Thus, our HOV method provides an efficient and reliable strategy for the routine cryopreservation of mouse embryos in animal facilities and biomedical laboratories, and for easy and cheap transportation.
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DOI：
10.1530/jrf.0.0890091
发表时间：
1990-05-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
KASAI, M;KOMI, JH;MACHIDA, T
通讯作者：
MACHIDA, T
影响因子：
2.8
作者：
Han, MS;Niwa, K;Kasai, M
通讯作者：
Kasai, M
影响因子：
2.8
作者：
HOCHI, S;FUJIMOTO, T;OGURI, N
通讯作者：
OGURI, N
影响因子：
2.8
作者：
Yavin, S.;Arav, A.
通讯作者：
Arav, A.
影响因子：
2.8
作者：
Martínez, AG;Matkovic, M
通讯作者：
Matkovic, M