动员外周血CD34阳性细胞流式细胞术测定中的策略探讨

Determination of CD34^+ Cells in Mobilized Peripheral Blood by Flow Cytometry

探索用流式细胞术测定动员外周血中CD34^+细胞的简便有效的方法,以便在干细胞动员中进行有效的动态监测,及时收取干细胞,把握最佳移植效果。将经药物动员的移植供者的外周血用CD34和CD45荧光抗体标记,用红细胞裂解液将红细胞裂解后经流式细胞仪检测。运用恰当的分析窗口、有效的设门分析结合干细胞的生物学特性,确定CD34^+细胞在窗口中的确切位置,统计CD34^+细胞在有核细胞中的比例。结果表明,通过本方法可有效地测定0.05 ％-0.1％的CD34^+细胞的相对含量,在药物动员的第5或6天,多数患者外周血中CD34^+细胞达到峰值,某些患者外周血中CD34^+细胞可超过1％。结论:通过标记中的有效阻断和多参数分析,本方法可对动员外周血中微量的CD34^+细胞进行有效的动态监测,对适时采集干细胞进行移植提供了重要的参考。

To explore a simple and effective method to determinate the volume of CD34 + cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry,the mobilized peripheral blood from donors and 100 fJ fresh whole blood were labeled with monoclonal antibodies anti-CD34-PE and anti-CD45-FITC,after lying the red blood cells,and assessed with flow cytometer FL2(log) vs SSC(log) and FLl(log) vs SSC(log) were mainly used for analysis windows.The results showed that a level of CD34+ cells in whole nucleated cells as low as 0.OS %-0.1% can be detected effectively using this method when 105 nucleated cells were counted.At day 5 or day 6,the level of CD34+ cells in most samples of patients reached a peak volume,some of samples and the levels were more than one percent in.It was concluded that CD34 cells can be effectively determined by using this method.According to the relative rate of CD34+ cells,the time to harvest the stem cells in blood can be determined.