ICE-FREE CRYOPRESERVATION OF MOUSE EMBRYOS AT -196-DEGREES-C BY VITRIFICATION
ICE-FREE CRYOPRESERVATION OF MOUSE EMBRYOS AT -196-DEGREES-C BY VITRIFICATION
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DOI：
10.1038/313573a0
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发表时间：
1985-01-01
期刊：
影响因子：
64.8
通讯作者：
FAHY, GM
中科院分区：
文献类型：
--
作者：
RALL, WF;FAHY, GM
复杂的哺乳动物器官（如肾脏）在冷冻至低温后无法发挥功能，被认为主要是由于细胞外冰形成对细胞间结构的机械破坏1 -5。通过施加超快速冷却和升温速率6 - 8或在冷却至−80 ° C期间逐渐降低平衡冰点9 - 13来避免冰形成的经典方法是不够的。另一种方法14 - 16依赖于高浓度的防冻剂水溶液过冷到非常低的温度的能力。在足够低的温度下，这些溶液变得如此粘稠，以至于它们固化而不形成冰，这一过程称为玻璃化。当使用常规程序冷冻保存胚胎悬浮液时，这种过冷行为允许细胞内玻璃化，即使在存在细胞外冰的情况下17 -20。因此，我们使用小鼠胚胎来研究在细胞内和细胞外溶液的玻璃化冷冻后获得高存活率的可行性，并在此报告，在适当控制的条件下，胚胎似乎在无冰冷冻保存后以高比例存活。
The failure of complex mammalian organs, such as the kidney, to function following freezing to low temperatures is thought to be due largely to mechanical disruption of the intercellular architecture by the formation of extracellular ice1–5. Classical approaches to the avoidance of ice formation through the imposition of ultra-rapid cooling and warming rates6–8or by gradual depression of the equilibrium freezing point during cooling to −80 °C9–13have not been adequate. An alternative approach14–16relies on the ability of highly concentrated aqueous solutions of cryoprotective agents to supercool to very low temperatures. At sufficiently low temperatures, these solutions become so viscous that they solidify without the formation of ice, a process termed vitrification. When embryo suspensions are cryopreserved using conventional procedures, this supercooling behaviour allows intracellular vitrification, even in the presence of extracellular ice17–20. We have therefore used mouse embryos to examine the feasibility of obtaining high survival following vitrification of both the intra- and extracellular solutions and report here that in properly controlled conditions embryos seem to survive in high proportions after cryopreservation in the absence of ice.