磁珠双阳性分选法在阵发性睡眠性血红蛋白尿症患者骨髓CD34^+CD59^+细胞体外扩增的应用  

Application of Double Immunomagnetic Positive Sorting to Ex Vivo Expansion of Marrow CD34^+CD59^+ Cells from Patients with Paroxysmal Nocturnal Hemoglobinuria

作　　者：肖娟[1] 武永吉[1] 张之南[1] 吕照江[1] 陈实平[2] 董红燕[2] 

机构地区：[1]中国医学科学院,中国协和医科大学北京协和医院血液科,北京100730 [2]中国医学科学院,中国协和医科大学基础医学研究所,北京100005

出　　处：《中国实验血液学杂志》2003年第2期179-183,共5页Journal of Experimental Hematology

基　　金：卫生部临床学科重点资金资助项目编号970 10 2 0 1

摘　　要：应用磁珠双阳性分选法在体外扩增阵发性睡眠性血红蛋白尿症 (PNH)患者的CD34+ CD5 9+ 细胞 ,为实现PNH患者进行临床自体骨髓移植 (ABMT)或自体外周血干细胞移植 (APBSCT)奠定基础。流式细胞术分选虽然经常用于细胞纯化 ,但难于满足大规模临床细胞分选的需要。本研究利用免疫磁珠系统 ,应用隔夜孵育法去除第一次分选时CD34+ 细胞吸附的磁珠 ,经过两次磁珠双阳性分选 ,从PNH患者骨髓中分离出足够数量的CD34+CD5 9+ 细胞 ,用于体外培养扩增。结果显示 :磁珠双阳性法分选的CD34+ CD5 9+ 细胞与磁珠 流式细胞术二步分选法比较 ,在细胞生存、增殖、扩增及形成造血集落形成单位 (CFU )的能力上均无显著性差异。结论 :磁珠双阳性分选法可能推广应用于其它双阳性或多阳性细胞的分离纯化 ,尤其是造血干Since flow cytometry was not feasible for sorting a huge amount of cells for clinical use, the method of double immunomagnetic positive sorting was used for selection of CD34 +CD59 + cells from bone marrow mononuclear cells in patients with paroxysmal nocturnal hemoglobinuria(PNH), which laid the groundwork for clinical ABMT/APBSCT of patients with PNH. Immunomagnetic positive selection was used for two times, the microbeads were removed from the CD34 + cells selected firstly by means of overnight culture, then the sufficient CD34 +CD59 + cells were used for ex vivo expansion. The results showed that the survival, proliferation and colony forming units of the selected CD34 +CD59 + cells by double immunomagnetic positive sorting had no significant difference as compared with that of CD34 +CD59 + cells selected by flow cytometry technique. It is suggested that the double immunomagnetic positive sorting promotes the use for separation and purification hematopoietic stem/progenitor cells and other cells with double or multiple markers cells for autologous hematopoietic stem cell transplantation in PNH patients.

关 键 词：磁珠双阳性分选法 阵发性睡眠性血红蛋白尿症 骨髓移植 干细胞移植 

分 类 号：R556.64[医药卫生—血液循环系统疾病] R457.7[医药卫生—内科学]