小鼠脾脏自然杀伤性树突细胞纯化方法的比较  

Comparison of methods for natural killer-dendritic cell sorting and purification from mouse spleen

作　　者：陈蓉明[1] 郑秋红[1] 龚福生[1] 应敏刚[2] 谢云青[1] 

机构地区：[1]福建省肿瘤医院肿瘤外科研究室,福建福州350014 [2]福建省肿瘤医院腹部外科,福建福州350014

出　　处：《实用肿瘤杂志》2012年第4期362-365,共4页Journal of Practical Oncology

基　　金：福建省卫生厅青年基金项目(2009-1-22)

摘　　要：目的比较流式细胞分选技术(FACS)及免疫磁珠分选技术(MACS)对小鼠脾脏自然杀伤性树突细胞NKDC分选纯化的效率,为自然杀伤性树突细胞的深入研究创造基本条件。方法 制备小鼠脾脏单细胞悬液,细胞悬液经70μm细胞滤膜过滤,离心滤液,用红细胞裂解液裂解沉淀中的红细胞,用缓冲液洗涤两次后通过FACS和MACS分选方法各自分选其脾脏表型为NK1.1+CD11c+的自然杀伤性树突细胞NKDC。结果 分选之前小鼠脾脏NK1.1+CD11c+自然杀伤性树突细胞占3.3%。单细胞悬液经FACS分选的NKDC纯度为(90.2±2.0)%,回收率为(91.3±1.8)%,细胞活力(82.5±1.2)%;经MACS分选纯度为(70.5±3.0)%,回收率为(80.5±1.7)%,细胞活力为(60.2±0.8)%。两种方法分选NKDC细胞纯度差异有统计学意义(P=0.03),回收率差异也有统计学意义(P=0.02)。结论 FACS较MACS可更高效、快捷、简便地纯化小鼠脾脏自然杀伤性树突细胞。Objective To compare the effectiveness of fluorescence-activated cell sorting （FACS） and magnetic activated cell sorting （MACS） in purification of natural killer-dendritic cells （NKDCs） from mouse spleen. Methods Spleens of male C57BL/6 （B6） mice were mechanically disrupted before being passed through a sterile 70 μm nylon mesh fiber. The resulting cell suspension was then pelleted （ 300 g x 7 min） , red ceils were lysed and the remaining cells were washed twice in media. Natural killer dendritic cells were sorted by FACS and MACS, respectively. Results Before sorting the proportion of the NKDCs in mouse spleens was 3.3 %. The purity of NKDCs after sorting by FACS and MACS reached to （ 90.2 ± 2.0） % and （70.5 ± 3.0） % , respectively （ P = 0.03,P 〈 0. 05） ; the recovery for FACS and MACS was （91.3± 1.8 ） % and （ 80.5 ± 1.7 ） %, respectively （ P = 0.02,P 〈 0. 05 ） ; the cell viability for FACS and MACS was （ 82.5 ± 1.2 ） % and （ 60.2 ±0.8 ） % , respectively. Conclusion FACS is more effective and efficient than MACS in sorting and purification of natural killer-dendritic cells from mouse spleen.

关 键 词：脾/细胞学 树突细胞 细胞分离 免疫磁化分离 流式细胞术 

分 类 号：R392[医药卫生—免疫学]