成人与胎儿骨髓间充质干细胞免疫特性比较

Adult and fetal bone marrow mesenchymal stem cell immune feature comparison

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。

We compared biological and immunological properties of aduh and fetal mesenchymal stem cells （MSCs） in order to provide experimental basis for experimental and clinical use. Adult and fetal normal bone marrow mononuclear cells were isolated by density gradiem eentrifugation and MSCs were obtained by adherent to plastic surface. MSC morphology was examined by inverted microscopy. MSC were idcntified by defining the surface markers on the cells of the 3rd passage using flow cytometry. Differentiation into fat ceils and osteoblasts was observed. Cytokines in the culture supernatant of cells of the 3rd generation were assayed by ELISA. MSCs of the 3rd generation were cocuhured with peripheral blood T cells to observe its effect on T cell proliferation. MTT was carried out to devel-op growth curves of MSCs. The resuhs showed that both the aduh and fetal bonc marrow derived MSCs were able to amplificate in vitro, and had similar morphology and phenotypes. Compared with fetal MSC, adult MSCs were stronger with respect to the capability of inhibiting T cells. Altbough both the duh arid fetal bone marrow derived MSC had immunosuppressive functions, overall, aduh MSCs were superior to the letus MSCs ill nmhiple aspects. Therefore, these findings suggesl thai the adult MSCs maybe more suitable than the fetal MSCs to be used as seed cells for tissue engineering as well as immunosuppressive agents. However, as the fetal MSCs are much easier available, and proliferated well in vitro, if nccessary, they can be used as an ideal substitute for adult MSC .