Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells
Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells
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DOI：
10.1002/bit.26880
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发表时间：
2019-03-01
影响因子：
3.8
通讯作者：
Hubel, Allison
中科院分区：
文献类型：
--
作者：
Pi, Chia-Hsing;Yu, Guanglin;Hubel, Allison
本研究检查了在海藻糖、蔗糖、甘油、甘露醇和肌酸等五种渗透剂的三种组合中冷冻保存的 Jurkat 细胞解冻后的恢复情况。使用低温拉曼光谱表征细胞反应，并使用统计模型分析解冻后恢复的变化。渗透剂的组合显示出解冻后恢复的明显趋势，并且观察到组合物与解冻后恢复之间的非线性关系，这表明不仅不同溶质之间存在相互作用，而且溶质与细胞之间也存在相互作用。在 1 ℃/min 的冷却速率下，优化的冷冻保护剂在不同渗透剂组合中的解冻后恢复与用 10% 二甲基亚砜测量的结果相当。统计模型用于了解各个渗透剂的重要性以及渗透剂之间的相互作用对解冻后恢复的影响。发现较高浓度的甘油以及糖和甘油之间的某些相互作用通常会增加解冻后的恢复。拉曼图像显示了渗透剂和渗透剂组合对冰晶形状的影响，反映了渗透剂与水之间的相互作用。成分的差异也会影响细胞内冰形成的存在或不存在，这也可以通过拉曼检测。这些研究帮助我们了解冷冻保护剂在这些渗透剂溶液中的作用方式。
This study examined the post-thaw recovery of Jurkat cells cryopreserved in three combinations of five osmolytes including trehalose, sucrose, glycerol, mannitol, and creatine. Cellular response was characterized using low-temperature Raman spectroscopy, and variation of post-thaw recovery was analyzed using statistical modeling. Combinations of osmolytes displayed distinct trends of post-thaw recovery, and a nonlinear relationship between compositions and post-thaw recovery was observed, suggesting interactions not only between different solutes but also between solutes and cells. The post-thaw recovery for optimized cryoprotectants in different combinations of osmolytes at a cooling rate of 1 degrees C/min was comparable to that measured with 10% dimethyl sulfoxide. Statistical modeling was used to understand the importance of individual osmolytes as well as interactions between osmolytes on post-thaw recovery. Both higher concentrations of glycerol and certain interactions between sugars and glycerol were found to typically increase the post-thaw recovery. Raman images showed the influence of osmolytes and combinations of osmolytes on ice crystal shape, which reflected the interactions between osmolytes and water. Differences in the composition also influenced the presence or absence of intracellular ice formation, which could also be detected by Raman. These studies help us understand the modes of action for cryoprotective agents in these osmolyte solutions.