人红细胞冻干程序优化的实验研究  

Optimization of lyophilization procedures for freeze-drying of human red blood cells

作　　者：陈麟凤[1] 刘景汉[1] 汪德清[1] 欧阳锡林[1] 庄远[1] 车辑[1] 于洋[1] 李卉[1] 

机构地区：[1]中国人民解放军总医院输血科,北京100853

出　　处：《南方医科大学学报》2010年第9期2055-2058,2062,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(30471931)~~

摘　　要：目的探讨冻干程序中几种参数设置对红细胞冻干复水后回收率的影响。方法将红细胞装入细管分别用四种模式对其进行降温,降温完成后将细管取投入25℃水浴中,对其外形变化进行比较判断玻璃化的程度;将冻好的样品放入冻干机搁板上,温度分别设置为-40、-50、-60、-70和-80℃,对样品进行冻干;对干燥开始不同温度、升温速率、红细胞样品在每一温度干燥的时间等多因素加以比较;最后测定了红细胞残余含水量。结果直接将样品投入液氮中降温,保护液在冷冻或溶化过程中形成玻璃化而未出现冰晶。搁板温度为-70℃和-80℃时红细胞回收率没有显著性差异(P>0.05),这两组与其他各温度下的回收率均有显著性差异(P<0.01)。E程序下的红细胞和血红蛋白回收率最高,分别为83.14%±9.55%和85.33%±11.42%。与其他各程序间存在显著差异(与A、B、C相比P<0.01;与D相比P<0.05)。随着残余含水量的增多,细胞回收率呈正相关的增加趋势。结论冻干中采用液氮快速降温,搁板温度预冷至-70℃,开始干燥时的温度越接近样品最初在搁板的平衡温度,红细胞升华干燥阶段升温速率越均匀缓和,红细胞的冻干效果越好。Objective To investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells（RBCs）.Methods Human RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 ℃.The morphological changes of the RBCs were observed to assess the degree of vitrification,and the specimens were placed in the freeze-dryer with the temperature set up at 40,-50,-60,-70 and-80 ℃.The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared,and the water residue in the specimens was determined.Results The protectant did not show ice crystal in the course of freezing and thawing.No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of-70 ℃ and-80 ℃（P〉0.05）.The E procedure resulted in the maximum recovery of the RBCs（83.14%±9.55%） and Hb（85.33%± 11.42%）,showing significant differences from the other groups（P〈0.01 or 0.05）.The recovery of the RBCs showed a positive correlation to the waterresidue in the samples.Conclusion Fast cooling in liquid nitrogen and shelf precooling at-70 ℃ with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.

关 键 词：冻干保护剂 冻干程序 搁板温度 残余含水量 

分 类 号：R318.52[医药卫生—生物医学工程]