改良法在制备冰冻红细胞技术上的应用研究  

An Improved Methods in the Preparation of Frozen Red Blood Cells

作　　者：刘慧 卓海龙 王道琳 陈婷婷 骆群 LIU Hui;ZHUO Hailong;WANG Daolin(The Fifth Medical Center of the General Hospital of the People's Liberation Army,Beijing 100071)

机构地区：[1]解放军总医院第五医学中心输血医学科,北京100071

出　　处：《临床输血与检验》2023年第3期315-319,共5页Journal of Clinical Transfusion and Laboratory Medicine

摘　　要：目的改良制备冰冻红细胞的甘油化过程,并对洗涤后的产品进行质量检测及分析评价。方法随机抽取本站4℃保存7 d内红细胞悬液(1 U)标本20份,随机分为两组,实验组在甘油化过程中,用等渗于生理盐水的0.25 mol/L的甘氨酸溶液,将57%的复方甘油试剂稀释至40%的甘油溶液,将甘油浓度降低,其他过程不变,对照组在甘油化过程中加入57%复方甘油试剂,将两组制备成冰冻红细胞,置于–80℃冰箱内保存,保存1个月后取出解冻去甘油化,比较两组冰冻解冻去甘油红细胞的质量指标,并进行分析评价。结果两组的游离血红蛋白,甘油残余量等指标均符合《全血及成分血质量要求(GB18469—2012)》,且实验组的游离血红蛋白(0.60±0.05)、甘油残余量(0.56±0.23)、红细胞内的甘油总量(0.82±0.05)、溶血率(0.33±0.10)与对照组相比显著下降,有统计学差异(P<0.05),实验组的三磷酸腺苷ATP(5.34±0.35)、2,3-DPG(538.40±59.96)与对照组相比显著升高,有统计学差异(P<0.05),实验组的红细胞回收率(86.9±2.18)与对照组相比没有明显差异(P>0.05),实验组的红细胞渗透脆性实验,开始溶血时的氯化钠浓度(4.28±0.33)与对照组相比有显著性差异(P<0.05),完全溶血时的氯化钠浓度(3.08±0.27)与对照组相比没有明显差异(P>0.05)。结论在制备冰冻红细胞的甘油化过程中,用甘氨酸溶液稀释后的40%浓度的甘油作为冰冻保护剂制备的冰冻红细胞,保存1个月后解冻去甘油后的红细胞产品与对照组相比,甘油残余量、红细胞内甘油总量、上清游离血红蛋白含量与溶血率都显著降低,三磷酸腺苷ATP与2,3-DPG水平显著升高,提高了冰冻红细胞的产品质量。Objective The glycerolization process for preparing frozen red blood cells(RBCs)was improved,and washed RBCs were tested and analyzed.Methods Twenty samples of suspended red blood cells(SRBCs)(one unit)stored at 4℃for 7 days were randomly divided into two groups.In the process of glycerolization,57%compound glycerol reagent was diluted to 40%glycerol solution with 0.25 mol/L glycine solution in isotonic saline,and the glycerol concentration was reduced,while other processes remained unchanged in the experimental group;57%compound glycerol reagent was added in the control group.The frozen RBCs prepared in the two groups were stored at–80℃for 1 month,then,were thawed for glycerolization.The quality indexes of frozen and thawed/deglycerolized RBCs(DRBCs)in the two groups were compared and analyzed.Results The indicators of free hemoglobin and residual glycerol in both groups were in line with the Quality Requirements for Whole Blood and Component Blood(GB 18469-2012).Free hemoglobin(0.60±0.05),residual glycerol(0.56±0.23),total glycerol in DRBCs(0.82±0.05),and hemolysis rate(0.33±0.10)in the experimental group were significantly decreased compared with the control group,with statistical difference(P<0.05).The ATP(5.34±0.35)and 2,3-DPG(538.40±59.96)in the experimental group were significantly higher than those in the control group(P<0.05).The recovery rate of DRBCs in the experimental group(86.9±2.18)was not significant difference compared with the control group(P>0.05).The osmotic fragility test of erythrocytes in the experimental group showed the sodium chloride concentration at the beginning of hemolysis(4.28±0.33)was significant difference(P<0.05),while the sodium chloride concentration at the complete hemolysis(3.08±0.27)was not significant difference(P>0.05).Conclusion During the glycerolization process of preparing frozen red blood cells,DRBCs prepared by using 40%glycerol diluted with glycine solution as cryoprotectants were stored for 1 month.Compared with the control group,the residual glycer

关 键 词：甘氨酸 甘油化过程 冰冻红细胞 

分 类 号：R331.141[医药卫生—人体生理学]