Spieles G, Kresin M, Loges K, Sputtek A, Heschel I, Rau G
Helmholtz-Institut für Biomedizinische Technik an der RWTH Aachen, Germany.
Cryobiology. 1995 Aug;32(4):366-78. doi: 10.1006/cryo.1995.1037.
A systematic study on the stability of frozen erythrocytes was performed. Washed and concentrated erythrocytes were mixed with an equal volume of cryoprotective solution containing 24% (w/w) hydroxyethyl starch (HES) and 60 mmol/liter NaCl according to an established protocol. Volumes of 250 microliters of this mixture were filled into polypropylene tubes and cooled to -196 degrees C with a rate of 293 degrees C/min by immersion in liquid nitrogen. The storage temperature was then varied from -10 to -75 degrees C and could be identified as the predominant factor influencing hemolysis kinetics. The effect of storage temperature on the frozen erythrocytes after thawing was evaluated by measuring the hemolysis in a dilute, isotonic NaCl solution (saline stability). A strong time dependence was found within the temperature range studied and could be described by an exponential kinetic law. A stability prediction was made for storage temperatures lower than those examined. Temperature ranges of qualitatively different hemolysis kinetics were identified and compared to devitrification behavior of intra-and extracellular solutions. The intracellular solution was simulated by a concentrated mixture of dried erythrocytes and water. The devitrification behavior was studied using DSC techniques. A rapidly frozen mixture was annealed at selected temperatures which fall into the range of storage temperatures for frozen erythrocytes. This paper tentatively interprets the devitrification data with respect to the means for cell damage during storage. The results are reviewed with respect to the design of a safe storage procedure.
对冷冻红细胞的稳定性进行了系统研究。按照既定方案，将洗涤并浓缩后的红细胞与等体积含24%（w/w）羟乙基淀粉（HES）和60 mmol/L氯化钠的冷冻保护溶液混合。将250微升该混合物装入聚丙烯管中，通过浸入液氮以293℃/分钟的速率冷却至-196℃。然后将储存温度在-10至-75℃之间变化，可确定其为影响溶血动力学的主要因素。通过测量在稀释的等渗氯化钠溶液中的溶血情况（盐水稳定性）来评估储存温度对解冻后冷冻红细胞的影响。在所研究的温度范围内发现了强烈的时间依赖性，可用指数动力学定律描述。对低于所检测温度的储存温度进行了稳定性预测。确定了溶血动力学性质不同的温度范围，并与细胞内、外溶液的脱玻化行为进行了比较。细胞内溶液用干燥红细胞和水的浓缩混合物模拟。使用差示扫描量热法（DSC）技术研究脱玻化行为。将快速冷冻的混合物在选定温度下进行退火处理，这些温度属于冷冻红细胞的储存温度范围。本文就储存期间细胞损伤的方式对脱玻化数据进行了初步解释。针对安全储存程序的设计对结果进行了综述。
