骨髓基质细胞的体外培养和成骨方向的诱导分化

Study on culture of bone marrow stromal cell in vitro and its osteogenic potential

目的:体外分离狗的骨髓基质细胞,诱导其成骨分化并鉴定成骨活性。方法:体外分离培养狗的骨髓基质细胞,传代培养中加入10-8mol/L地塞米松、50μg/ml维生素C和10mmol/Lβ-甘油磷酸钠进行诱导分化,进行细胞形态和增殖观察,矿化结节Von Kossa染色,细胞碱性磷酸酶染色和Ⅰ型胶原免疫组化染色检测其成骨活性。结果:骨髓基质细胞经诱导后表现出明显的成骨活性,体外矿化结节Von Kossa钙染色阳性;传代细胞碱性磷酸酶染色阳性,酶活性>80%,Ⅰ型胶原免疫组化染色强阳性。结论:体外分离培养的骨髓基质细胞中含有骨源性前体细胞,传代细胞具有较强的成骨潜能。

Objective:To testify whether bone marrow stromal cells(BMSCs) could be differentiated into osteoblasts in vitro and identify the osteogenic potential of the cells. Methods: BMSCs isolated from the Mongrel dogs were cultured in vitro. 50μg/ml ascorbic acid ,10mmol/L Na-β-Glycerophosphate and 10-8mol/L dexamethasone were added in the subculture medium as the inducers. The morphological characters of BMSCs and cells proliferation were obsevered with phase contrast microscope. Alkaline phosphatase activities of the sub culturing cells was detected. Mineralized nodules in the culture dishes were stained by Von Kossa histochemical methods, and typeⅠcollagen was detected by immunohistochemical staining.Results:Cultured BMSCs showed the ability of osteogenic potential and could be differentiated into osteoblasts in RPMI1640 essential medium. Von Kossa staining of mineralized nodules were positive. Alkaline phosphatase staining and immunohistochemical staining of typeⅠcollagen were both positive in the passage cells. Conclusions: BMSCs cultured in vitro contained osteogenic precursor cells and had powerful osteogenic potential.