骨髓基质细胞-成骨细胞复合培养体系的建立

Construction of a co-culture system for human bone marrow stromal cells-osteoblasts in vitro

背景:在骨修复和重建过程中,成骨细胞和骨髓基质细胞是主要功能细胞,二者存在着密切的功能联系。目的:通过骨髓基质细胞-成骨细胞共育体系的建立,观察共育体系中两种细胞之间的功能影响及生物学特点。方法:原代分离人骨髓基质细胞和人成骨细胞,将2种细胞置于Transwell共育环境中共同培养,建立人骨髓基质细胞-成骨细胞共育体系。分别采用MTT、丫啶橙染色、碱性磷酸酶活性检测等方法初步评价共育体系中两种细胞增殖、凋亡及功能改变情况。结果与结论:在复合培养体系中,骨髓基质细胞-成骨细胞共育体系能够促进成骨细胞的增殖与碱性磷酸酶的活性,同时抑制骨髓基质细胞的凋亡,促进骨髓基质细胞的趋化聚集。结果提示在复合培养体系中,骨髓基质细胞能够加速成骨细胞的增殖及成骨活性,另外成骨细胞也可减少骨髓基质细胞的凋亡,并加强其成骨性分化的作用。两者之间具有较为紧密的影响和功能联系。

BACKGROUND： Bone marrow stromal cells and osteoblasts playa major role during bone repair and reconstruction and the two are closely associated with each other functionally. OBJECTIVE： To establish a co-culture system for bone marrow stromal cells-osteoblasts in vitro so as to investigate the features of both kinds of cells in one system.METHODS： Human bone marrow stromal cells and osteoblastswere primary isolated and these two kinds of cells were co-cultured in a Transwell system to establish a co-culture system for human bone marrow stromal cells-osteoblasts. The proliferation, apoptosis and other features of human bone marrow stromal cells and osteoblasts were determined by MTT assay, acridine orange staining and alkaline phosphatase activities. RESULTS AND CONCLUSION： The co-culture system for human bone marrow stromal cells and osteoblasts promoted the proliferation of osteoblasts, increased alkaline phosphatase expression, inhibited apoptosis of bone marrow stromal cells and promoted the aggregation of bone marrow stremal cells. These results indicate that in the co-culture system for human bone marrow stromal cells and osteoblasts, bone marrow stromal cells can increase the osteogenic ability of osteoblasts by accelerating their proliferation; in addition, osteoblasts can lower the apoptosis of bone marrow stromal cells and strengthen their osteogenic differentiation. These findings suggest a close functional association between human bone marrow stromal cells and osteoblasts.