小容量红细胞的冰冻保存:血型抗原反应性评估及其应用价值  

Cryopreservation of small-volume red blood cells:evaluation of blood group antigen reactivity and its application value

作　　者：赵亚玲 王艳侠[1] 王子夜 马斯禹 邵伟 张媛媛[1] 江欣[1] 甘佳[1] ZHAO Yaling;WANG Yanxia;WANG Ziye;MA Siyu;SHAO Wei;ZHANG Yuanyuan;JIANG Xin;GAN Jia(Department of Blood Transfusion,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]中国医学科学院北京协和医院输血科,北京100730

出　　处：《中国输血杂志》2025年第3期352-357,387,共7页Chinese Journal of Blood Transfusion

基　　金：中央高水平临床科研专项项目(2022-PUMCH-B-057)。

摘　　要：目的 建立小容量(≤1 mL)红细胞冰冻保存方案,评估红细胞冰冻后的血型抗原反应性和稳定性,探索其在免疫血液学参考实验室中的应用价值。方法 小容量红细胞冰冻保存120 d后,通过解冻、去甘油等步骤恢复红细胞成分,对红细胞的质量进行评估;血清抗体梯度稀释后分别与冰冻前后的红细胞反应,记录凝集评分,进而定量评估冰冻前后Rh、Duffy、Lewis、Kidd、M、H等血型抗原的反应性及稳定性;采用流式细胞术分析冰冻前后红细胞的ABO抗原表达百分比及平均荧光强度,以评估冰冻红细胞在流式免疫分析及血型亚型研究中的可用性。结果 解冻去甘油后的红细胞溶血率为(0.27±0.10)%,上清游离血红蛋白(0.52±0.14)g/L,红细胞回收率为(69.12±7.91)%;解冻后的红细胞DAT均为阴性,冰冻前后红细胞血型抗原的反应性无差异,冰冻前后红细胞表面A抗原及B抗原表达百分比及平均荧光强度无差异。结论 小容量红细胞冰冻保存方案可应用于参考实验室的免疫血液学分析,有望在血型鉴定、抗体筛查、鉴定及血型相关的研究中广泛应用。Objective To establish a cryopreservation protocol for small-volume(≤1 mL)red blood cells(RBCs)and to evaluate the reactivity and stability of blood group antigens after cryopreservation,so as to explore its potential appli-cation in immunohematology reference laboratories.Methods Small-volume RBCs were cryopreserved for 120 days,fol-lowed by thawing and deglycerolization to restore the RBC components.The quality of the RBCs was assessed.Serum anti-bodies were serially diluted and reacted with RBCs before and after cryopreservation,and agglutination scores were recorded to quantitatively evaluate the reactivity and stability of blood group antigens such as Rh,Duffy,Lewis,Kidd,M,and H.Flow cytometry was used to analyze the percentage and mean fluorescence intensity of ABO antigen expression on RBCs be-fore and after cryopreservation to assess the usability of cryopreserved RBCs in flow immunophenotyping and blood group subtype studies.Results The hemolysis rate of thawed and deglycerolized RBCs was(0.27±0.10)%,with a supernatant free hemoglobin level of(0.52±0.14)g/L,and the RBC recovery rate was(69.12±7.91)%.The direct antiglobulin test(DAT)was negative for all thawed RBCs.There was no difference in the reactivity of blood group antigens before and after cryopreservation,and no difference in the percentage and mean fluorescence intensity of A and B antigen expression on RBCs before and after cryopreservation.Conclusion The small-volume RBC cryopreservation protocol can be applied to immunohematology analysis in reference laboratories and is expected to be widely used in blood group identification,antibody screening,identification,and blood group-related research.

关 键 词：冰冻红细胞 小容量红细胞保存 血型抗原反应性 流式分析 免疫血液学 

分 类 号：R457.11[医药卫生—治疗学] R331.141[医药卫生—临床医学]