直接接触共培养技术在体外探索细胞间相互作用的应用研究

发布时间：2018-09-08 21:52

【摘要】：目的:建立观察直接接触共培养体系中单一种类细胞生长变化的研究方法,探索直接接触共培养技术在体外细胞之间相互作用研究中的应用。方法:大鼠软骨细胞与荧光标记的大鼠骨髓间充质干细胞以1∶2的比例平面共培养7天。共培养2天时,应用共聚焦显微镜观测共培养体系中两种细胞的增殖情况;共培养7天时,应用流式分选技术分选出单一种类细胞,然后分析其成软骨分化特异性基因的表达。结果:经过共培养后,骨髓间充质干细胞比例减少;共培养组中软骨细胞增殖率高于单独培养组;在共培养体系中骨髓间充质干细胞的成软骨分化趋势明显,但软骨细胞成软骨分化特异性基因表达下降。结论:本实验应用流式细胞术以及共聚焦显微镜技术成功检测了直接接触共培养体系中骨髓间充质干细胞和软骨细胞各自的细胞增殖与成软骨分化特异性基因的表达,明确了直接接触共培养技术可以有效地应用于体外细胞间相互作用的研究。
[Abstract]:Aim: to establish a method to observe the growth and change of single cell in direct contact co-culture system, and to explore the application of direct contact co-culture in the study of cell interaction in vitro. Methods: rat chondrocytes and fluorescent labeled rat bone marrow mesenchymal stem cells were co-cultured at 1:2 level for 7 days. The proliferation of two kinds of cells in co-culture system was observed by confocal microscope for 2 days, and the expression of specific genes for chondrogenic differentiation was analyzed by flow sorting technique after 7 days of co-culture. Results: after co-culture, the proportion of bone marrow mesenchymal stem cells decreased, the proliferation rate of chondrocytes in co-culture group was higher than that in single culture group, and the trend of chondrogenic differentiation of bone marrow mesenchymal stem cells in co-culture system was obvious. However, the specific gene expression of chondroblast differentiation was decreased. Conclusion: flow cytometry and confocal microscopy were used to detect the proliferation of bone marrow mesenchymal stem cells and chondrocytes respectively and the expression of specific genes for chondrogenic differentiation of bone marrow mesenchymal stem cells and chondrocytes. It is clear that direct contact co-culture can be used effectively in the study of inter-cell interaction in vitro.
【作者单位】：北京大学第三医院运动医学研究所;北京大学医学部生物化学与分子生物学系;
【基金】：国家自然科学基金项目(81171726)
【分类号】：R329

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