Clark P, Hawkins H E, Karow A M
Cryobiology. 1985 Apr;22(2):156-60. doi: 10.1016/0011-2240(85)90169-5.
Renal cortical slices were frozen to various subzero temperatures after treatment with 2.1 M of one of three cryoprotectants, dimethyl sulfoxide (Me2SO), ethylene glycol, or glycerol. The effects on tissue [K+]/[Na+] of cooling to these temperatures were tested (using identical procedure times, cooling rates, and warming rates) by holding the slices at each experimental temperature for appropriate periods of time prior to rewarming. The effects of the holding time were assessed by comparison with slices which were cooled and rewarmed with no intermediate holding time. Slices treated with ethylene glycol or glycerol were found to exhibit a continuous decrease in [K+]/[Na+] with lowered temperatures, in contrast to those treated with Me2SO. Slices treated with Me2SO actually experienced a continuous increase in [K+]/[Na+] with lowered temperature (-12 to -33 degrees C). Me2SO does exhibit toxic effects at subzero temperatures. Adverse effects of holding time on viability are seen for Me2SO-treated slices at higher subzero temperatures. These effects were alleviated as the temperature is reduced, suggesting that temperature has a greater effect on survival of renal cortical tissue than Me2SO concentration. However, the toxicity observed at higher subzero temperatures is expected to be of importance, particularly for slowly cooled tissues which are exposed to these temperatures for relatively long periods of time.
肾皮质切片在用三种冷冻保护剂（二甲基亚砜（Me2SO）、乙二醇或甘油）中的一种2.1M处理后，被冷冻至不同的零下温度。通过在复温前将切片在每个实验温度下保持适当时间（使用相同的程序时间、冷却速率和升温速率），测试冷却至这些温度对组织[K+]/[Na+]的影响。通过与无中间保持时间直接冷却和复温的切片进行比较，评估保持时间的影响。发现用乙二醇或甘油处理的切片随着温度降低[K+]/[Na+]持续下降，这与用Me2SO处理的切片相反。用Me2SO处理的切片实际上随着温度降低（-12至-33摄氏度）[K+]/[Na+]持续增加。Me2SO在零下温度确实表现出毒性作用。在较高的零下温度下，观察到保持时间对用Me2SO处理的切片的活力有不利影响。随着温度降低，这些影响得到缓解，这表明温度对肾皮质组织存活的影响比Me2SO浓度更大。然而，在较高的零下温度下观察到的毒性预计很重要，特别是对于缓慢冷却且在这些温度下暴露相对较长时间的组织。
