NUCLEATION AND GROWTH OF ICE CRYSTALS INSIDE CULTURED-HEPATOCYTES DURING FREEZING IN THE PRESENCE OF DIMETHYL-SULFOXIDE
NUCLEATION AND GROWTH OF ICE CRYSTALS INSIDE CULTURED-HEPATOCYTES DURING FREEZING IN THE PRESENCE OF DIMETHYL-SULFOXIDE
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DOI：
10.1016/s0006-3495(93)81319-5
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发表时间：
1993-12-01
影响因子：
3.4
通讯作者：
TONER, M
中科院分区：
文献类型：
--
作者：
KARLSSON, JOM;CRAVALHO, EG;TONER, M
一个三部分，耦合模型的细胞脱水，成核，晶体生长被用来研究细胞内冰形成（IIF）在培养的肝细胞冷冻在二甲基亚砜（DMSO）的存在下。非均质成核温度作为DMSO浓度的函数进行预测，并与实验数据吻合良好。模拟冷冻方案正确地预测和解释了实验观察到的冷却速率、升温速率和储存温度对肝细胞功能的影响。对于冷却至-40 ℃的细胞，冷却速率小于10 ℃/min时不会发生IIF。IIF确实在更快的冷却速率下发生，并且预测的细胞内冰的体积随着冷却速率的增加而增加。以5 ℃/min冷却至-80 ℃的细胞显示在-46.8 ℃下经历成核，结果是高于该值的储存温度导致与升温速率无关的高存活率，而较低的储存温度导致缓慢升温速率的细胞损伤。细胞损伤与预测的细胞内冰量呈正相关，临界冰含量的上限估计为等渗水含量的3.7%。该模型的力量是有限的，在低温下作为DMSO浓度的函数，在估计的细胞溶质粘度和膜渗透性的困难。
A three-part, coupled model of cell dehydration, nucleation, and crystal growth was used to study intracellular ice formation (IIF) in cultured hepatocytes frozen in the presence of dimethyl sulfoxide (DMSO). Heterogeneous nucleation temperatures were predicted as a function of DMSO concentration and were in good agreement with experimental data. Simulated freezing protocols correctly predicted and explained experimentally observed effects of cooling rate, warming rate, and storage temperature on hepatocyte function. For cells cooled to -40-degrees-C, no IIF occurred for cooling rates less than 10-degrees-C/min. IIF did occur at faster cooling rates, and the predicted volume of intracellular ice increased with increasing cooling rate. Cells cooled at 5-degrees-C/min to -80-degrees-C were shown to undergo nucleation at -46.8-degrees-C, with the consequence that storage temperatures above this value resulted in high viability independent of warming rate, whereas colder storage temperatures resulted in cell injury for slow warming rates. Cell damage correlated positively with predicted intracellular ice volume, and an upper limit for the critical ice content was estimated to be 3.7% of the isotonic water content. The power of the model was limited by difficulties in estimating the cytosol viscosity and membrane permeability as functions of DMSO concentration at low temperatures.