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比较成人与胎儿骨髓间充质干细胞(MSC)免疫学特性的差异,为实验室及临床MSC的应用提供依据。采用密度梯度离心法分离正常成人和胎儿骨髓单个核细胞,通过贴壁法培养骨髓MSC。以倒置显微镜观察细胞形态;取第3代细胞以流式细胞术进行表面标志鉴定;观察向脂肪细胞及成骨细胞诱导分化情况;收集第3代细胞培养上清,用ELISA法检测上清液中细胞因子水平;将第3代细胞与外周T细胞共培养,观察其对T细胞增殖的影响,用MTT法绘制MSC生长曲线。研究发现,成人与胎儿来源的骨髓MSC都具有体外扩增能力,并且形态相似、表型相同。成人MSC对T细胞的抑制能力强于胎儿MSC。成人及胎儿来源的骨髓MSC都具有免疫抑制功能,而且,成人MSC在免疫调节功能上优于胎儿MSC,因此更适合于作组织工程的种子细胞及免疫抑制剂。
To compare the immunological characteristics of adult and fetal bone marrow mesenchymal stem cells (MSCs), and to provide a basis for the application of laboratory and clinical MSC. Normal adult and fetal bone marrow mononuclear cells were isolated by density gradient centrifugation, and bone marrow MSCs were cultured by adherent method. The morphology of the cells was observed with an inverted microscope. The surface markers of the third generation of cells were identified by flow cytometry. Differentiation into adipocytes and osteoblasts was observed. The third generation of cells were harvested and supernatants The third generation cells were co-cultured with peripheral T cells to observe their effects on the proliferation of T cells. The growth curve of MSCs was drawn by MTT method. The study found that adult and fetal bone marrow derived MSC have the ability to expand in vitro, and the morphological similar, the same phenotype. Adult MSCs inhibit T cells more strongly than fetal MSCs. Adult and fetal bone marrow MSCs are immunosuppressive and adult MSCs are superior to fetal MSCs in immune regulation and are therefore more suitable for tissue engineering of seed cells and immunosuppressive agents.