人骨髓间充质干细胞对脐血T淋巴细胞转化的影响

Effect of Human Bone Marrow Mesenchymal Stem Cell on Cord Blood T Lymphocyte Transformation

为研究间充质干细胞 (mesenchymalstemcell,MSC)的免疫调节作用 ,从人骨髓中分离培养间充质干细胞 ,并通过其形态的均一性及流式细胞术检测其表面标志以鉴定其纯度。将分离培养的间充质干细胞接种于 96孔板中 (分别为 2 0 0 0个 孔组与 10 0 0个 孔组 ) ,与经分离的脐血T淋巴细胞共培养 3天 ,以单独培养的脐血T淋巴细胞为对照组 ,加入植物血凝素 (PHA)刺激 6 0小时 ,H3 TdR标记后用液体闪烁计数仪检测 ,以观察MSC对脐血T淋巴细胞转化的影响。结果显示 ,当接种 2 0 0 0个MSC时 ,其对T细胞的增殖起抑制作用 (抑制率为 33.4 % ) ;而接种 10 0 0个MSC时 ,其对T细胞的增殖起促进作用 (促进率为 2 2 .5 % )。由此得出结论 ,MSC免疫调控作用与其加入细胞数量相关 ,当MSC数量多时表现为负调控 。

To study the effect of mesenchymal stem cell (MSC) on immune function, MSCs were isolated and cultured from human bone marrow cells. The purity of MSCs were identified with the spindle fibroblastic morphology characterization by microphotograph and the phenotypes were tested by flow cytometry. MSCs were plated in 96 well plates (2 000/well and 1 000/well), and cocultured for 3 days with T cells isolated from cord blood. Cord blood T cells non cocultured with MSC acted as control group. After cord blood T cells stimulated by PHA for 60 hours, thymidine was added to each well and T cell proliferation was assessed by thymidine incorporation. The results showed that cord blood T cell proliferation was suppressed when 2 000 MSCs were plated each well and cord blood T cell proliferation was activated when 1 000 MSCs were plated. Our results suggested that the immunomodulatory function of MSC seemed dependent on cell dose. High concentration of MSC most often resulted in inhibition, while low concentration resulted in stimulation.