Three-dimensional collagen gel culture promotes osteoblastic phenotype in bone marrow derived cells.
Three-dimensional collagen gel culture promotes osteoblastic phenotype in bone marrow derived cells.
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三维胶原凝胶培养促进骨髓来源细胞的成骨细胞表型。
DOI：
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
K. Mizuno
中科院分区：
文献类型：
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作者：
Seiji Kinoshita;Maureen A Finnegan;R. Bucholz;K. Mizuno
当颅细胞或成骨细胞系在I型胶原凝胶中培养时，与单层培养相比，可以观察到早期和广泛的钙化。骨髓源性细胞在三维I型胶原凝胶中培养，观察细胞能否向成骨细胞分化。在有效诱导成骨分化方面，我们通过形态学、碱性磷酸酶活性、I型胶原和骨桥蛋白mRNA表达，将胶原凝胶培养与I型胶原包被培养皿培养和无胶原塑料培养皿培养进行了比较。骨髓原代细胞形成由阳性表达碱性磷酸酶活性的成纤维细胞组成的菌落。在胶原凝胶中培养的原代和第三代细胞均观察到矿物沉积，而在塑料培养皿中培养的第三代细胞没有矿化。胶原凝胶中的细胞碱性磷酸酶活性高于其他两种方法，表明三维胶原网络有效地促进了成骨细胞的分化。胶原凝胶中I型胶原mRNA的表达量在第3代细胞中提高了3倍，在原代细胞中略有降低。胶原凝胶中骨桥蛋白mRNA的表达量在第3代细胞培养中提高4倍，而在原代细胞培养中降低。这些结果表明，胶原凝胶培养可能是诱导骨髓传代细胞成骨的有益环境。
When calvarial cells or osteogenic cell lines were cultured in type I collagen gel, calcification was observed early and diffusely compared to monolayer culture. Bone marrow derived cells were cultured in three-dimensional type I collagen gel to investigate whether the cells can differentiate into osteogenic cells. In terms of efficient induction of osteogenic differentiation, we compared collagen gel culture to type I collagen coated dish culture and collagen-free plastic dish culture by morphology, alkaline phosphatase activity, and mRNA expression for type I collagen and osteopontin. Bone marrow derived primary cells formed colonies consisting of fibroblastic cells positively expressing alkaline phosphatase activity. Mineral deposition was observed in both primary and the 3rd passaged cells cultured in collagen gel, whereas the 3rd passaged cells on plastic dishes failed to be mineralized. Cells in collagen gel showed higher alkaline phosphatase activity than those in the other two methods suggesting that three-dimensional collagen network stimulated osteoblastic differentiation effectively. The expression level for type I collagen mRNA of the cells in collagen gel was three times higher in the 3rd passaged cells, and was slightly decreased in primary cells compared to the other two methods. The osteopontin mRNA expression of the cells in collagen gel was four times higher in the 3rd passaged cell culture but lower in primary cell cultures. These results suggested that collagen gel culture might be a useful environment for osteogenic induction of passaged cells derived from bone marrow.