不同标记法及去红细胞法对检测微量脐血CD34^+造血干细胞含量的影响  

The effect of different labeling and red cell removed methods on measuring the CD34^+ cells of the cord blood

作　　者：范华骅[1] 仇志根[1] 郑滨[1] 袁纪军[1] 高峰[1] 

机构地区：[1]上海市血液中心,200051

出　　处：《临床输血与检验》2003年第1期7-9,共3页Journal of Clinical Transfusion and Laboratory Medicine

摘　　要：目的　采用 ISHAGE(international society of hematotherapy and graft engineering)法和常用的 CD34+造血干细胞流式检测方法对同一脐血样品进行比较实验 ,确定更适用于微量脐血 CD34+造血干细胞含量检测的方法。方法　分别采用 ISHAGE造血干细胞计数协会推荐方法、低渗 NH4 Cl去红细胞 CD4 5 / CD34双色标记法 (NH4 Cl双标法 )、Optilyse C溶血剂去红细胞 CD34单色标记法 (Optilyse C单标法 )、低渗 NH4 Cl去红细胞 CD34单色标记法 (NH4 Cl单标法 )、羟乙基淀粉沉淀去红细胞 CD4 5 / CD34双色标记法 (HES双标法 )检测脐血造血干细胞含量 ,并比较不同方法测得数据的差异。结果　 8份脐血标本用 ISHAGE方法测得 CD34+造血干细胞含量的中位数为 0 .2 78% ,NH4 Cl双标法测得结果为 0 .2 97% ,与 ISHAGE方法相比差异无显著性。用 HES双标、Optilyse C单标法和 NH4 Cl单标法测得的结果分别为 5 .715 % ,0 .391%和 0 .74 1% ,与ISHAGE方法相比差异有显著性。结论　 ISHAGE方法是一种公认的准确性高 ,重复性好的检测造血干细胞方法。Optilyse C单标法、NH4 Cl单标法和 HES双标法测定结果比实际值偏高。Objective\ The ISHAGE and some other common methods of measuring CD34 + cells with flow cytometry were used to determine the same sample of cord blood to confirm which method was more adequate.\ Method\ The ISHAGE,Optilyse C single fluorescence labeling,HES double fluorescence labeling,NH 4Cl single fluorescence labeling and NH 4Cl double fluorescence labeling methods were used to determine the CD34 + concentration of the cord blood.\ Results\ The medium of CD34 + concentration of 8 cord blood samples using the ISHAGE method was 0 278%.The result using the NH 4Cl double fluorescence labeling method was 0 297%,which had no significant difference with the ISHAGE method.The results using the HES double fluorescence labeling,Optilyse C single fluorescence labeling and NH 4Cl single fluorescence labeling were 5 715%,0 391% and 0 741%,respectively.They had significant difference with the ISHAGE method.Conclusion ISHAGE method is an acknowledged one that is accurate and precise to determine the concentration of hematopoietic cells.The results of the Optilyse C single fluorescence labeling,NH 4Cl single fluorescence labeling and HES double fluorescence labeling methods are higher than the actual count.

关 键 词：CD34^+造血干细胞 脐血 含量 检测方法 ISHAGE法 流式细胞仪检测法 OptilyseC单标法 NH4C1单标法 HES双标法 标记法 

分 类 号：R446.11[医药卫生—诊断学]