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目的研究体外培养大鼠骨髓基质干细胞的方法及其生长特点和成骨能力。方法通过贴壁培养法和密度梯度离心法分离成年大鼠骨髓基质干细胞,应用含维生素C、地塞米松、β甘油磷酸钠的诱导分化培养液定向诱导传代细胞向成骨细胞分化,检测碱性磷酸酶活性和细胞矿化作用。结果原代培养基质干细胞首先形成细胞集落,14d时集落间接近融合;传代细胞体积变大,约5~7d传代一次。诱导条件下,细胞碱性磷酸酶活性明显增高,并出现了矿化结节。结论骨髓基质干细胞易于体外分离培养及扩增,成骨能力较强,可作为骨组织工程的种子细胞。
Objective To study the method of culturing rat bone marrow stromal cells in vitro and its growth characteristics and osteogenesis ability. Methods Adult rat bone marrow stromal cells (MSCs) were isolated by adherent culture and density gradient centrifugation. Differentiated osteoblasts were differentiated into osteoblasts by inducing differentiation medium containing vitamin C, dexamethasone and β-glycerophosphate. Phosphatase activity and cell mineralization. Results Primary culture of MSCs first formed colonies of cells, and the colonies approached and fused on the 14th day. The passage cells became larger and were passaged once every 5 to 7 days. Under the induction conditions, the cell alkaline phosphatase activity was significantly increased, and the emergence of mineralized nodules. Conclusion BMSCs can be easily isolated, cultured and expanded in vitro with strong osteogenic potential and can be used as seed cells for bone tissue engineering.