免疫磁性活化细胞分选方法优化及纯化后细胞的生物学特性检测  

Optimization of magnetic activated cell sorting and the biological characteristics of isolated cells

作　　者：耿珊[1] 张琛[1] 刘俊[1] 刘典锋[1] 周玥[1] 王亚平[1] 

机构地区：[1]重庆医科大学干细胞与组织工程研究室,组织胚胎学教研室,重庆400016

出　　处：《细胞与分子免疫学杂志》2012年第7期752-754,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81173398)

摘　　要：目的:改良常规免疫磁性活化细胞分选(MACS)的分离纯化方法,提高分选后细胞的纯度并确保细胞仍具有良好活性及功能。方法:以干细胞抗原-1(Sca-1)标记的Sca-1+造血干/祖细胞(Sca-1+HSC/HPC)为例,分别用改良后和常规MACS法分选出小鼠骨髓细胞Sca-1+细胞,流式细胞术检测两种分选方法获得Sca-1+细胞纯度;计算阳性细胞回收率;台酚蓝染色检测分选细胞存活百分率;CCK-8检测细胞增殖,混合造血祖细胞(CFU-Mix)体外培养评价分选Sca-1+细胞分化能力。结果:改良MACS法分选获得的Sca-1+细胞纯度达93%以上(常规MACS法仅为87%),阳性细胞的回收率可达73%(常规法仅为62.3%);细胞存活率、细胞增殖和CFU-Mix集落形成能力两种分选方法无明显差别。结论:改良法能明显提高细胞分选纯度及回收率,细胞活性和功能保持良好,值得各种细胞免疫磁性分选参考采用。AIM： To optimize the traditional magnetic activated cell sorting （MACS） so as to enhance the purification and keep the viability of ceils after separation. METH- ODS： Take stem cell antigen-1 labeled hemopoietic stem/ progenitor cells （Sca-1^＋ HSC/HPC） for example. The tradi- tional MACS and the optimized MACS were applied to obtain the Sca-1^＋ HSC/HPC from mouse bone marrow respectively. The purifications of Sca-1^＋ cells from two groups were tested by flow cytometry; The survival rates of Sca-1^＋ cells from two groups were detected by trypan blue dye; The proliferation of Sca-1^＋ cells was detected by cell counting kit-8 （CCK-8） ; the differentiation capacity of Sca-1^＋ cells was measured by CFU-Mix. RESULTS： The purification of Sca-1^＋ cells was up to 93% by the optimized MACS compared to 87% by the traditional one; the recovery rate of positive cells was 73% by the optimized MACS compared to 62.3% by the traditional one; there was no statistical difference in the viability and proliferation of Sca-1^＋ cells and the capacity of producing CFU-Mix between two groups. CON- CLUSION= The optimized MACS can significantly enhance the purification and recovery rate of Sca-1^＋ cells without influence of cell viability and biological features, which can be considered to apply in other MACS.

关 键 词：免疫磁性活化细胞分选 方法 干细胞抗原-1 生物学特征 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]