树突状细胞外排体促进间充质干细胞体外增殖  

Dendritic cell-derived exosomes promote in vitro proliferation of mesenchymal stem cells

作　　者：王姿[1] 陈津[2] 林珊珊[2] 郑晓丽[1] 郭子宽[2] 王恒湘[1] 闫洪敏[1] 

机构地区：[1]空军总医院血液科,北京100036 [2]军事医学科学院放射与辐射医学研究所实验血液研究室

出　　处：《临床血液学杂志》2014年第4期589-592,共4页Journal of Clinical Hematology

摘　　要：目的：探讨树突状细胞（DC）外排体（DCex）对人骨髓间充质干细胞（MSC）体外增殖的影响。方法：利用人骨髓单个核细胞诱导DC,超速离心法从细胞培养上清中分离提取DCex,采用透射电镜观察所提取的DCex表征形态,并通过流式细胞技术检测其表面抗原以确定其来源。MSC培养体系中分别加入不同蛋白质浓度的DCex,培养24、48、72h后MTT法检测MSC增殖状态,并且利用CFSE标记及流式细胞学分析进一步验证DCex对MSC的增殖作用。结果：人骨髓DC具有典型的形态学和表型特征。透射电镜下DCex呈典型的圆形或椭圆形膜层结构,直径为40-100nm。DCex表达DC细胞的标志物。MTT实验增殖结果显示DCex具有促进MSC增殖作用,且其效应呈浓度和时间依赖性。CFSE标记流式结果也证实DCex对MSC的促增殖作用。结论：DCex可以促进MSC体外增殖,可能与其携带母细胞成分而成为细胞间信息交流的新方式有关,但是具体机制尚需进一步阐明。Objective：To investigate the role of the dendritic cell-derived exosome（DCex）on the proliferation of bone mesenchymal stem cells（MSC）.Method：DCs were prepared from human bone marrow mononucleated cells.The culture supernatants were collected and DCex were harvested by ultracentrifugation.The morphology of DCex was observed under a transmission electron microscope and the phenotypes of DCex were detected by flow cytometry.The test was divided into 6groups in line with DCex concentration（0,1,2,5,10and 15μg·ml^-1）.The DCex were added into the culture of MSCs and the cell proliferation was tested by MTT test and a membrane dye dilution assay.Result：Under transmission electron microscope,the DCex were either spherical or oval membrane vesicles with diameters of about 40to 100nm.The DCex expressed surface molecules specific for DCs.DCex promoted MSC proliferation in dose-and time-dependent manners.The growth-promoting effect was also identified by the CFSE dilution assay.Conclusion：DCex can promote the in vitro proliferation of MSC,though the underlying mechanisms need further clarified.

关 键 词：树突状细胞 外排体 间充质干细胞 增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]