人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究  

The Process of Coculturing Peripheral Blood Mononuclear Cells and Human Umbilical Cord Derived Mesenchymal Stem Cells Observed with Atomic Force Microscope

作　　者：龚超群[1] 蔡继业[1] 郜世隽[2] 王秋兰[1] 金花[1] 

机构地区：[1]暨南大学生命科学技术学院化学系,广州510632 [2]暨南大学医学院微生物学免疫学教研室,广州510632

出　　处：《中国生物工程杂志》2010年第8期6-12,共7页China Biotechnology

基　　金：海外及港澳学者合作研究基金(30828028);国家自然科学基金(30872404)资助项目

摘　　要：采用原子力显微镜与倒置显微镜在细胞层次上观察了人外周单个核细胞(PBMCs)与同种异源脐带间充质干细胞(hUC-MSCs)共培养的过程,并在单细胞水平上分析了共培养前后人外周单个核细胞的形貌和生物物理性质。结果发现:共培养后贴壁人外周单个核细胞的形态发生了很大的改变,并且表面分布着大小不一的颗粒状聚合物。利用AFM高空间分辨的力位移曲线测量系统,发现共培养72h后培养上清中人外周单个核细胞、贴壁的人外周单个核细胞的粘滞力分别是单纯培养72h的人外周单个核细胞的2倍、5倍,而细胞的硬度分别是单纯培养人外周单个核细胞的1.5倍、2倍。CCK-8检测提示,共培养过程中,干细胞的生长与外周血单个核细胞的生长出现了竞争作用。通过AFM探测人外周单个核细胞与脐带间充质干细胞共培养的可视化数据,有助于更好地了解间充质干细胞与外周血单个核细胞的相互作用。The inverted microscope was used to observe the process that peripheral blood mononuclearcells （PBMCs） were cocuhured with allogeneic human umbilical cord derived mesenchymal stem cells （ hUC- MSCs）. The atomic force microscope was used to analyze the morphology and mechanical property after two kinds of cells were cocultured. The result showed that the morphology of adherence PBMCs changed greatly after coeultured, and was coated with different grain sizes of polymers. From the high spatial resolution of AFM force- curves, the adhesion force and stiffness values of PBMCs coeultured with hUC-MSCs were stronger than those of controlled PBMCs. CCK-8 indicated that PBMCs may appear to compete with hUC- MSCs in the co-culture system. The visualized data obtained from AFM can provide a better understanding the interactive mechanism of hUC- MSCs and PBMCs.

关 键 词：脐带间充质干细胞 外周血单个核细胞 原子力显微镜 倒置荧光显微镜 

分 类 号：Q819[生物学—生物工程]