Armitage W J, Juss B K, Easty D L
Department of Ophthalmology, University of Bristol, Bristol Eye Hospital, United Kingdom.
Cryobiology. 1995 Feb;32(1):52-9. doi: 10.1006/cryo.1995.1004.
The effects of 30 min exposure to 10% v/v cryoprotectant (glycerol, dimethyl sulfoxide, or propane-1,2-diol) on intercellular junctions were investigated in monolayers of epithelial (MDCK) cells. The cells were grown on microporous membrane filters and transepithelial resistance was monitored as an index of junction integrity. At 22 degrees C, Me2SO had the least effect of the three cryoprotectants, causing a gradual fall in transepithelial resistance of only 17% in 30 min. This compares with falls of 50% in 15 min and 37% over 30 min, respectively, with glycerol and propane-1,2-diol. At 0 degrees C, however, propane-1,2-diol had no effect on transepithelial resistance, whereas the effect of Me2SO was now similar to that of glycerol, viz. a sustained reduction in resistance of approximately 30% throughout the 30-min exposure. The results could not be explained solely in terms of osmotic effects and suggested that the cryoprotectants were directly affecting components of the junctional complex or associated elements of the cytoskeleton. During removal of the cryoprotectants by two-step dilution, changes in transepithelial resistance showed little consistency between the different cryoprotectants. This suggested that the pattern of change was dictated more by the state of the junctions immediately before removal of the cryoprotectants, with each cryoprotectant having affected the junctions to different extents and perhaps by different mechanisms, than by the removal of cryoprotectant itself. Thus, intercellular junctions in epithelial sheets were affected by even modest concentrations of cryoprotectant, but the effect was influenced by the type of cryoprotectant and by temperature of exposure.(ABSTRACT TRUNCATED AT 250 WORDS)
在单层上皮（MDCK）细胞中研究了暴露于10%（体积/体积）冷冻保护剂（甘油、二甲基亚砜或1,2 - 丙二醇）30分钟对细胞间连接的影响。细胞生长在微孔膜滤器上，监测跨上皮电阻作为连接完整性的指标。在22摄氏度时，三种冷冻保护剂中二甲基亚砜的影响最小，30分钟内跨上皮电阻仅逐渐下降17%。相比之下，甘油和1,2 - 丙二醇在15分钟内分别下降50%，30分钟内下降37%。然而，在0摄氏度时，1,2 - 丙二醇对跨上皮电阻没有影响，而此时二甲基亚砜的影响与甘油相似，即在整个30分钟暴露期间电阻持续降低约30%。结果不能仅用渗透效应来解释，表明冷冻保护剂直接影响连接复合体的成分或细胞骨架的相关元件。在通过两步稀释去除冷冻保护剂的过程中，不同冷冻保护剂之间跨上皮电阻的变化几乎没有一致性。这表明变化模式更多地取决于去除冷冻保护剂之前连接的状态，每种冷冻保护剂对连接的影响程度不同，可能机制也不同，而不是取决于冷冻保护剂本身的去除。因此，上皮片层中的细胞间连接即使受到适度浓度冷冻保护剂的影响，但这种影响受冷冻保护剂类型和暴露温度的影响。（摘要截断于250字）
