Department of Chemistry, University of Ottawa, D'Iorio Hall, 10 Marie Curie, Ottawa, ON, Canada K1N 6N5.
Glycobiology. 2012 Jan;22(1):123-33. doi: 10.1093/glycob/cwr115. Epub 2011 Aug 17.
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.
各种单糖和二糖的冰晶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相关。将细胞用二甲基亚砜 (DMSO) 冷冻保存的对照实验与冷冻保存后的细胞存活率进行了比较。二糖的 220mM 溶液是最有效的冰晶重结晶抑制剂；然而，当使用 200mM 的 D-半乳糖溶液时，获得了最佳的细胞活力。该溶液在生理温度下的细胞毒性最小，并且在冷冻-解冻过程中有效地保存了细胞。事实上，这种碳水化合物与 5%DMSO 溶液一样有效。进一步的研究表明，D-半乳糖的保护作用是由于其内化及其减轻渗透应激、防止细胞内冰形成和/或抑制冰晶重结晶的能力。本研究支持这样的假设，即冷冻保护剂抑制冰晶重结晶的能力是提高冷冻后细胞活力的一个重要特性。这种冷冻保护作用在三种不同的人细胞系中都有观察到。此外，我们证明了潜在冷冻保护剂抑制冰晶重结晶的能力可以用作其在亚低温下保存细胞能力的预测指标。
