人脐带间充质干细胞对再生障碍性贫血患者T细胞相关因子调节的体外研究

目的体外观察人脐带间充质干细胞对再生障碍性贫血患者和正常对照T细胞相关细胞因子IL-2、IFN-γ、IL-17、IL-6的调节作用.方法采用组织块培养法从脐带分离、培养间充质干细胞.用流式细胞术鉴定脐带间充质干细胞的表型.分离再生障碍性贫血患者和正常对照外周血单个核细胞与脐带间充质干细胞共培养.用ELISA法测定外周血单个核细胞组和共培养组培养上清中细胞因子IL-2、IFN-γ、IL-17和IL-6的水平.结果脐带来源间充质干细胞高表达CD29、CD44、CD105、HLA-I.人脐带间充质干细胞与再生障碍性贫血患者外周血单个核细胞共培养后上清中IL-2水平无明显变化（P〉0.05）而IFN-γ水平有所下降（P〉0.05）,IL-17水平和IL-6水平明显上升（P〈0.01）.人脐带间充质干细胞与正常对照外周血单个核细胞共培养后上清中IL-2和IFN-γ水平明显下降（P〈0.05）而IL-17水平无明显变化（P〉0.05）,IL-6水平明显上升（P〈0.01）.结论人脐带间充质干细胞明显可抑制正常外周血单个核细胞分泌IL-2、IFN-γ,其与外周血单个核细胞共培养后可升高IL-6水平.

Regulatory Effects of Human Uumbilical Cord-mesenchymal Stem Cells on Cytokines Related to T Cells in vitro in Aplastic Anemia Patients

Objective To explore the regulatory effects of human umbilical cord-mesenchymal stem cells（hUC-MSCs）on cytokines（IL-2,IFN-γ,IL-17,and IL-6）related to T cell in aplastic anemia patients and normals in vitro.Methods MSCs were isolated from human umbilical cord and cultured by tissue culture method,and the phenotype of MSCs was indentificated by flow cytometry.Mononuclearcells were isolated from peripheral blood of patients with aplastic anemia and normals,and then were cocultured with hUC-MSCs.The levels of IL-2,IFN-γ,IL-17,and IL-6 in supernatants of coculture were detected by enzyme-linked immunosorbent assay.Results CD29,CD44,CD105,and HLA-I were highly expressed in hUC-MSCs.After coculture of hUC-MSCs and mononuclearcells of aplastic anemia patients,the levels of IL-2 in culture supernatants had no significant changes（P0.05）,the levels of IFN-γdecreased a little,but the difference had no statistical significance（P0.05）,the levels of IL-17 and IL-6 increased significantly（P0.01）.After coculture of hUC-MSCs and mononuclearcells of normal controls,the levels of IL-2 in culture supernatants decreased（P0.05）,the levels of IFN-γdecreased significantly（P0.05）,the levels of IL-17 had no significant changes（P0.05）,the levels of IL-6 increased significantly（P0.01）.Conclusions hUC-MSCs can inhibit the secretion of IL-2 and IFN-γby mononuclearcells of normal controls.The levels of IL-6 in culture supernatants increase significantly after coculture of hUC-MSCs and mononuclearcells.