大鼠骨髓间充质干细胞骨向分化及体外诱导实验

An experimental study of osteogenic differentiation and inducted in vitro of SD rat bone marrow stem cell

目的建立骨髓间充质干细胞(MSCs)体外分离培养体系,进行骨向分化诱导,并将诱导的成骨细胞复合钛网体外成骨,为MSCs进一步的临床应用研究提供实验依据。方法用密度梯度离心法分离大鼠骨髓MSCs,并对其形态学特征进行观察。用诱导剂对MSCs向成骨细胞进行诱导分化,并进行形态学观察和免疫细胞化学检测。用扫描电镜观察骨向诱导后细胞复合钛网的情况。结果用密度梯度离心法成功分离获得了高纯度的骨髓MSCs。经骨向诱导后,ALP和矿化结节染色阳性。扫描电镜可观察到骨向诱导后细胞复合在钛网表面有矿化的基质沉积。结论成功建立了大鼠骨髓MSCs体外分离和培养体系,MSCs能够向成骨细胞分化,骨向诱导后细胞复合钛网体外有矿化的基质。

Objective To establish an isolation and culture system of rat bone marrow stem cells（MSCs）, osteogenic differentiation and compound titanium meshes in vitro in order to verify their osteogenesis, Methods Density gradient centrifugation was used to isolate bone MSCs from SD rats and their morphological characteristics were examined under microscope. The bone MSCs underwent osteogenic induction and cytochemical and immunocytochemical staining were performed to verify their multipotential. Cells of osteogenic differentiation that compounded titanium meshes were detected by scanning electron microscope. Results High purity of bone MSCs was successfully obtained and they showed good proliferation ability. Both ALP activity and mineraliztion node staining were positive in bone MSCs after osteogenic differentiation. Cells of osteogenic differentiation that compounded titanium meshes were detected by scanning electron microscope, it was deposited calcified matrix. Conclusion The isolation and culture system of rat bone MSCs was successfulIy established and the bone MSCs sustained their osteogenic differentiation potential in vitro. The cells of osteogenic differentiation that compounded titanium meshes was deposited calcified matrix.