Principles of Ice-Free Cryopreservation by Vitrification
Principles of Ice-Free Cryopreservation by Vitrification
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DOI：
10.1007/978-1-0716-0783-1_2
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发表时间：
2021-01-01
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影响因子：
--
通讯作者：
Wowk, Brian
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文献类型：
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作者：
Fahy, Gregory M.;Wowk, Brian
玻璃化是冷冻保存的替代方法，其能够使水合活细胞在没有冰的情况下冷却至低温。玻璃化简化并经常改善冷冻保存，因为它消除了来自冰的机械损伤，消除了寻找最佳冷却和升温速率的需要，消除了混合细胞类型群体中细胞的不同最佳冷却和升温速率的重要性，消除了寻找溶液效应损伤和细胞内冰形成之间经常不完美的折衷的需要，并且可以通过使用快速冷却而使冷害“跑过”，而没有细胞内冰形成的风险。另一方面，玻璃化需要比通过冷冻的冷冻保存高得多的浓度的冷冻保护剂，这引入了更大的渗透损伤和冷冻保护剂毒性的风险。幸运的是，在过去的35年中，已经发现了大量针对后一个问题的补救措施，并且在大多数情况下，通过仔细注意冷冻保护剂的引入和冲洗技术，可以消除或充分控制渗透损伤。因此，玻璃化冷冻有可能使上级和方便的低温保存的生物系统（包括分子，细胞，组织，器官，甚至一些整个生物体）的范围广泛，它也越来越被认为是一个成功的策略，在自然界中的恶劣环境条件下生存。但是，玻璃化冷冻的潜力有时会受到对所涉及的复杂物理和生物学原理理解不足的限制，因此，更好的理解不仅有助于改善目前的结果，而且还可能为未来可能更成功的新策略指明方向。本章相应地描述了玻璃化冷冻的基本原理，并指出了这种替代冷冻保存方法的广泛潜在生物学意义。
Vitrification is an alternative to cryopreservation by freezing that enables hydrated living cells to be cooled to cryogenic temperatures in the absence of ice. Vitrification simplifies and frequently improves cryopreservation because it eliminates mechanical injury from ice, eliminates the need to find optimal cooling and warming rates, eliminates the importance of differing optimal cooling and warming rates for cells in mixed cell type populations, eliminates the need to find a frequently imperfect compromise between solution effects injury and intracellular ice formation, and can enable chilling injury to be "outrun" by using rapid cooling without a risk of intracellular ice formation. On the other hand, vitrification requires much higher concentrations of cryoprotectants than cryopreservation by freezing, which introduces greater risks of both osmotic damage and cryoprotectant toxicity. Fortunately, a large number of remedies for the latter problem have been discovered over the past 35 years, and osmotic damage can in most cases be eliminated or adequately controlled by paying careful attention to cryoprotectant introduction and washout techniques. Vitrification therefore has the potential to enable the superior and convenient cryopreservation of a wide range of biological systems (including molecules, cells, tissues, organs, and even some whole organisms), and it is also increasingly recognized as a successful strategy for surviving harsh environmental conditions in nature. But the potential of vitrification is sometimes limited by an insufficient understanding of the complex physical and biological principles involved, and therefore a better understanding may not only help to improve present outcomes but may also point the way to new strategies that may be yet more successful in the future. This chapter accordingly describes the basic principles of vitrification and indicates the broad potential biological relevance of this alternative method of cryopreservation.