大鼠骨髓基质干细胞体外培养及成骨作用的实验研究

目的研究大鼠骨髓基质干细胞在体外条件培养下的成骨能力，为骨组织工程选择理想的种子细胞来源。方法取大鼠骨髓基质干细胞进行体外培养，用倒置相差显微镜、钙染色、扫描电镜和X线能谱等手段观察细胞的生长情况和矿化能力。结果细胞呈贴壁型生长，形态为梭形或多角形，在条件培养液中这些细胞间不发生接触抑制，而是相互重叠成多层，形成钙化结节，与成骨细胞的特点相符合。碱性磷酸酶染色呈强阳性，结节钙染色阳性，X线能谱分析显示结节的主要组成元素为钙和磷。结论体外培养的大鼠骨髓基质干细胞具有与成骨细胞相似的形态特征，并能在体外形成钙化的新骨结节。

The Culture of Rat Bone Marrow Stromal Cells and Their Osteogenesis in Vitro

Objective To study osteogenic capability of rat marrow stromal cells in vitro and to find out an ideal source of seed cells for bone tissue engineering. Methods Bone marrow cells were isolated from four-week-old Sprague-Dawley rats and cultured in DMEM medium. The growth status of all cultured cells was observed continually on a phase-contrast microscope and a scanning electron microscope, and the capacity of mineralization was detected with calcifying-nodule staining and X-ray energy spectrum analysis. Results Cultured cells grew anchoring in fusiform or multi-angulation shape, and there were no contact inhibition in cells in conditioned medium. Cells grew in multi-layer and formed calcifying nodules in accordance with osteoblasts. ALP staining and mineralized nodules staining were positive. The main elements of nodules included calcium and phosphorus by X-ray energy spectrum analysis. Conclusion The cultured cells from rat bone marrow shows osteoblastic characteristics in shape and can form calcified nodule in vitro.