重症肌无力患者造血干/祖细胞及其亚群富集纯化的研究

Collection and purification of myasthenia gravis CD34 + hematopoictic stem (progenitor) cells and its sup-groups

目的:获得高纯度造血干/祖细胞及其亚群,用于重症肌无力(MG)造血干细胞移植治疗和生物学特性的研究。方法:用免疫磁珠法(Mini MACS)从MG骨髓中富集CD 34+细胞后再用荧光激活细胞分选(Fluorescent activatedcell sorter,rAGS)纯化CD34+/CD38+和T CD34+/CD38-细胞亚群。结果:MiniMACS分选的CD34+细胞群纯度达90％,FACS分选的CD34+/CD38-和CD34+/CD38+两细胞亚群纯度可达95％以上。结论:联合使用MiniMACS和FACS可迅速大量纯化HSC/HPC及其重要细胞亚群,可用于临床造血干细胞移植、基因治疗和研究HSC/HPC及其亚群生物学特性。

Objective: To collect and purify the bone marrow CD34 + hematopofetic stem celts and its sub-groups trom the patients with myasthenia gravis. Methods: CD34 + cells were collected and purified by means of imunomagnetic beads on MiniMACS device. Flow cytometry was emplyed to determine percentage of CD34 + cells, CD34 + /CD38 cells and CD34 + /CD38 + cells before and after miniMACS. And to detect cell ability after CD34 + cell purification, Results: The purity of CD34 + cells a-mount to more than 90% after, the purification. The purity of CD34 + /CD38- cells and CD34 + /CD38 + cells amount to more than 95% after FACS. Conclusions: The Conbination of MiniMACS and FACS can sort effectively the bone marrow CD34 + cells and its sub-groups so that the bone marrow CD34 + cells and its sub-groups are used for hematopoietic stem cell transplantation, gene therapy and for study of HSC/HPC and its cell sub-groups.