Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada.
Cryobiology. 2010 Aug;61(1):46-51. doi: 10.1016/j.cryobiol.2010.04.005. Epub 2010 May 21.
Cryopreservation plays a key role in the long-term storage of native and engineered cells and tissues for research and clinical applications. The survival of cells and tissues after freezing and thawing depends on the ability of the cells to withstand a variety of stresses imposed by the cryopreservation protocol. A better understanding of the nature and kinetics of cellular responses to temperature-induced conditions is required to minimize cryoinjury. An interrupted freezing procedure that allows dissection of cryoinjury was used to investigate the progressive damage that occurs to cells during cryopreservation using slow cooling. Simulations of cellular osmotic responses were used to provide interpretation linking states of the cell with events during the freezing procedure. Simulations of graded freezing (interrupted slow cooling without hold time) were correlated with cell recovery results of TF-1 cells. Calculated intracellular supercooling and osmolality, were used as indicators of the probability of cryoinjury due to intracellular ice formation and solution effects, providing direct links of cellular conditions to events in the freezing process. Using simulations, this study demonstrated that both intracellular supercooling and osmolality are necessary to explain graded freezing results.
冷冻保存在研究和临床应用中对天然和工程细胞和组织的长期储存起着关键作用。细胞和组织在冷冻和解冻后的存活取决于细胞承受冷冻保存方案施加的各种应激的能力。需要更好地了解细胞对温度诱导条件的性质和动力学反应，以最大程度地减少低温损伤。使用中断的冷冻程序允许在使用缓慢冷却进行冷冻保存时对细胞的渐进性损伤进行剖析。细胞渗透反应的模拟用于提供与冷冻过程中的事件相关的解释，将细胞状态与冷冻过程中的事件联系起来。分级冷冻（无暂停时间的中断缓慢冷却）的模拟与 TF-1 细胞的细胞恢复结果相关联。计算出的细胞内过冷和渗透压用作由于细胞内冰形成和溶液效应而导致低温损伤的概率的指标，为细胞状态与冷冻过程中的事件提供了直接联系。本研究通过模拟表明，细胞内过冷和渗透压都是解释分级冷冻结果所必需的。
