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目的研究原代培养的人肺成纤维母细胞体外分化特性。方法于2006年3月至2006年10月在清华大学第一附属医院中心实验室,将原代培养的人肺成纤维母细胞分别在成骨细胞培养基(含地塞米松,维生素C和β-磷酸甘油)和成脂肪细胞培养基(含马血清,地塞米松和胰岛素)作用下进行分化诱导。组织化学方法行碱性磷酸酶和钙化斑块染色,Westernblotting法测定骨桥蛋白表达,油红染色鉴定脂肪细胞形成。结果在成骨细胞培养基诱导下,细胞内碱性磷酸酶表达增加,于第14天可见大量钙盐沉积和骨桥蛋白表达增加。在成脂肪细胞培养基作用第14天,部分细胞内出现脂肪小滴聚集。结论人肺成纤维母细胞具有多分化潜能,能够在一定条件下向成骨细胞和脂肪细胞分化,表现出骨髓间充质干细胞的特性。
Objective To study the in vitro differentiation characteristics of primary cultured human lung fibroblasts. Methods From March 2006 to October 2006 at the Central Laboratory of the First Affiliated Hospital of Tsinghua University, primary cultured human lung fibroblasts were cultured in osteoblast medium (containing dexamethasone, vitamin C and β - phosphoglycerol) and adipogenic medium (containing horse serum, dexamethasone and insulin) induced differentiation. Histochemical staining was performed on alkaline phosphatase and calcified plaque. Osteopontin protein expression was determined by Western blotting and adipocyte formation by oil red staining. Results The expression of alkaline phosphatase increased in osteoblast medium, and a large amount of calcium deposition and osteopontin expression were observed on day 14. On the 14th day of adipocyte medium function, fat droplets accumulated in some cells. Conclusion Human lung fibroblasts have the potential of differentiation and can differentiate into osteoblasts and adipocytes under certain conditions and show the characteristics of bone marrow mesenchymal stem cells.