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

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NH4C1去红细胞CD45／CD34双色标记法(NH4C1双标法)、OptilyseC溶血剂去红细胞CD34单色标记法(OptilyseC单标法)、低渗NH4C1去红细胞CD34单色标记法(NH4C1单标法)、羟乙基淀粉沉淀去红细胞CD45／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0．278％，NH4C1双标法测得结果为0．297％，与ISHAGE方法相比差异无显性。用HES双标、OptilyseC单标法和NH4C1单标法测得的结果分别为5．715％，0．391％和0．741％，与ISHAGE方法相比差异有显性。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。OptilyseC单标法、NH4C1单标法和HES双标法测定结果比实际值偏高。

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

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.