间充质干细胞与巨噬细胞共培养体系的细胞因子表达模式研究  

The expression pattern of cytokines in mesenchymal stem cells and macrophages co-cultured system

作　　者：高弘斐 张潜[2] 陈龙 宋公宇 赵玉洁[1] 万卫红[1] 刘祖林[1] 余丽梅[1] 章涛[1] GAO Hongfei ZHANG Qian CHEN Long SONG Gongyu ZHAO Yujie WAN Weihong LIU Zulin YU Limei ZHANG Tao(Key Laboratory of Cell Engineering of Guizhou Province （Affiliated Hospital of Zunyi Medical College）, Zunyi 563003, China Department of Human Anatomy, Zunyi Medical College, Zunyi 563003, China Experimental Medical Center, Dongfeng Hospital Affiliated to Hubei University of Medicine, Shiyan 442000, China)

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,563003 [2]遵义医学院附属医院解剖学教研室,563003 [3]湖北医药学院附属东风医院实验中心,十堰442000

出　　处：《免疫学杂志》2017年第11期930-936,共7页Immunological Journal

基　　金：贵州省生物治疗人才基地建设项目(黔人领发[2013]5号)

摘　　要：目的比较外周血间充质干细胞(PBMSCs)和骨髓间充质干细胞(BMMSCs)与巨噬细胞(M0型)共培养后的细胞因子表达差异。方法分离培养大鼠PBMSCs、BMMSCs和M0型巨噬细胞,采用Transwell培养体系分别进行PBMSCs和BMMSCs与M0共培养,同时还采用PBMSCs、BMMSCs和M0单独培养组作为对照。共培养3 d后采用Bio-Plex免疫微球法对各组培养上清液中IL-10、IFN-γ、IL-1β、TNF-α细胞因子含量进行检测分析。结果与PBMSCs组比较,PBMSCs+M0共培养上清液中的IL-10、IL-1β含量升高(P<0.05),TNF-α的含量降低(P<0.05),而IFN-γ的含量没有变化(P>0.05);BMMSCs+M0共培养组的细胞因子表达变化模式同PBMSCs+M0共培养组,但变化程度均高于后者(P<0.05)。结论 PBMSCs和BMMSCs与M0共培养均可促进IL-10、IL-1β表达,抑制TNF-α的表达,且BMMSCs的调节能力强于PBMSCs,表明PBMSCs和BMMSCs均具有炎症免疫调节潜力并具有差异性,为间充质干细胞在不同疾病状态下的治疗应用提供了新的实验依据。To investigate the changes of cytokines expression in macrophages（M0） cocultured withblood-derived mesenchymal stem cells（PBMSCs） or bone marrow-derived mesenchymal stem cells（BMMSCs）, ratPBMSCs, BMMSCs and macrophages were isolated, cultured, and identified, and 5 groups were set as PBMSCs,BMMSCs, macrophages, PBMSCs co-cultured with macrophages（PBMSCs＋M0）, BMMSCs co-cultured withmacrophages（BMMSCs＋M0）. All cells were cultured for 3 days and then the supernatant was collected to detect theconcentration of cytokines IL-10, IFN-γ, IL-1β and TNF-α by Bio-Plex immune-microsphere assay. Macrophagesco-cultured with PBMSCs/BMMSCs showed significantly higher levels of IL-10 and IL-1β（P〈0.05）, lower level ofTNF-α in the supernatant（P〈0.05）, and unchanged level of IFN-γ（P〈0.05）, as compared with PBMSCs/BMMSCs.The changes of all cytokines in BMMSCs＋M0 group were more significant, as compared with PBMSCs＋M0 group（P〈0.05）. In conclusion, co-culture of PBMSCs/BMMSCs with macrophage can stimulate the secretion of IL-10 andIL-1β, reduce the secretion of TNF-α, and the inflammatory modifying ability of BMMSCs is stronger that that ofPBMSCs, which provides a new strategy for the application of mesenchymal stem cells in different disease states.

关 键 词：外周血 间充质干细胞 巨噬细胞 Transwell共培养 细胞因子 

分 类 号：R392[医药卫生—免疫学]