Inhibiting ice recrystallization and optimization of cell viability after cryopreservation
Inhibiting ice recrystallization and optimization of cell viability after cryopreservation
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DOI：
10.1093/glycob/cwr115
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发表时间：
2012-01-01
期刊：
影响因子：
4.3
通讯作者：
Ben, Robert N.
中科院分区：
文献类型：
--
作者：
Chaytor, Jennifer L.;Tokarew, Jacqueline M.;Ben, Robert N.
各种单糖和双糖的冰重结晶抑制活性与其在不同浓度下冷冻保存人细胞系的能力有关。将低温保存后的细胞存活率与二甲基亚砜（DMSO）低温保存的对照实验进行比较。最有效的冰再结晶抑制剂是220 mM的双糖溶液；然而，当使用200 mM的d-半乳糖溶液时，细胞存活率最高。该溶液在生理温度下具有最低的细胞毒性，并在冻融期间有效保存细胞。事实上，这种碳水化合物和5% DMSO溶液一样有效。进一步的研究表明，d-半乳糖的冷冻保护作用是由于其内化和减轻渗透胁迫、防止细胞内冰形成和/或抑制冰再结晶的能力。这项研究支持了冷冻保护剂抑制冰再结晶的能力是提高细胞冻融后活力的重要特性的假设。在三种不同的人类细胞系中观察到这种冷冻保护作用。此外，我们证明了一种潜在的冷冻保护剂抑制冰重结晶的能力可能被用作其在零下温度下保存细胞的能力的预测因子。
The ice recrystallization inhibition activity of various mono- and disaccharides has been correlated with their ability to cryopreserve human cell lines at various concentrations. Cell viabilities after cryopreservation were compared with control experiments where cells were cryopreserved with dimethylsulfoxide (DMSO). The most potent inhibitors of ice recrystallization were 220 mM solutions of disaccharides; however, the best cell viability was obtained when a 200 mM d-galactose solution was utilized. This solution was minimally cytotoxic at physiological temperature and effectively preserved cells during freeze-thaw. In fact, this carbohydrate was just as effective as a 5% DMSO solution. Further studies indicated that the cryoprotective benefit of d-galactose was a result of its internalization and its ability to mitigate osmotic stress, prevent intracellular ice formation and/or inhibit ice recrystallization. This study supports the hypothesis that the ability of a cryoprotectant to inhibit ice recrystallization is an important property to enhance cell viability post-freeze-thaw. This cryoprotective benefit is observed in three different human cell lines. Furthermore, we demonstrated that the ability of a potential cryoprotectant to inhibit ice recrystallation may be used as a predictor of its ability to preserve cells at subzero temperatures.