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。
Objective To investigate the application of direct contact co-culture technique in exploring cell interaction in vitro. Methods Rat chondrocytes were co-cultured with fluorescent labeled mesenchymal stem cells of bone marrow at 1：2 ratio in monolayer for 7 days. At day 2,observation of proliferation of single cell type was performed through confocal microscope in this co-culture system; and at day 7,the single cell type was separated by flow cytometry,then its chondrogenic marker genes expression profile was analyzed. Results After co-culture,the ratio of bone marrow mesenchymal stem cells decreased; the proliferation rate of chondrocytes in co-culture group was higher than mono-cuhure group; bone marrow mesenchymal stem cells showed a significant chondrogenic differentiation trend in co-culture system,while chondrocytes showed decreased chondrogenic marker genes expression. Conclusion This study successfully demonstrates that direct ＇contact co-culture technique can be effectively applied to investigate cell interaction in vitro.
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
2013年第4期337-342,352,共7页
Chinese Journal of Sports Medicine
基金
国家自然科学基金项目(81171726)
关键词
共培养
荧光标记
软骨细胞
骨髓间充质干细胞
co-culture ,fluorescent labeling, chondrocyte, bone marrow mesenchymal stem cell
