大鼠骨髓间充质干细胞的体外成骨诱导*

背景：骨髓间充质干细胞作为良好的种子细胞,将其复合于生物支架治疗骨缺损取得了良好的进展,是当今研究的一大热点。目的：观察大鼠骨髓间充质干细胞体外诱导向成骨样细胞分化的效果。方法：采用贴壁筛选法分离培养SD大鼠骨髓间充质干细胞,取生长状态良好的第3代骨髓间充质干细胞,分为2组,对照组细胞仅用DMEM/F12培养基培养;实验组以含成骨诱导剂的DMEM/F12培养基培养。结果与结论：全骨髓贴壁法培养的原代骨髓间充质干细胞呈梭形或多角形贴壁生长;经成骨诱导剂诱导后骨髓间充质干细胞呈圆形或卵圆形贴壁生长,碱性磷酸酶活性明显强于对照组(P < 0.01);茜素红染色出现阳性的钙化结节;Western blotting检测Ⅰ型胶原蛋白表达较对照组有明显增加(P < 0.01);骨钙素ELISA定量分析较对照组明显升高(P < 0.01)。提示全骨髓贴壁法培养骨髓间充质干细胞方法简单、实用,所培养的骨髓间充质干细胞在成骨诱导后表现了成骨细胞的形态学和生物学特性。

In vitro osteogenic induction of rat bone marrow mesenchymal stem cells

BACKGROUND: Bone marrow mesenchymal stem cells are a kind of good seed cells. Bone marrow mesenchymal stem cells compounded onto biological scaffold for treatment of bone defects have acquired satisfactory outcomes and become a gradually increasing research area. OBJECTIVE: To investigate the effect of in vitro osteogenic induction of rat bone marrow mesenchymal stem cells.METHODS: Sprague-Dawley rat bone marrow mesenchymal stem cells were in vitro isolated by adherent method. Passage 3 well growing bone marrow mesenchymal stem cells were divided into two groups. The control group was cultured only with DMEM/F12 culture medium. The experimental group was cultured with DMEM/F12 culture medium containing osteogenic inductor. RESULTS AND CONCLUSION: Primary bone marrow mesenchymal stem cells cultured by the whole bone marrow adherence method exhibited a shuttle-shaped or polygon-shaped appearance. After induced by osteogenic inductor, bone marrow mesenchymal stem cells adhered to plate wall in a circle or oval-shaped manner. Alkaline phosphatase activity was significantly higher in the experimental group than in the control group (P < 0.01). Calcified nodules appeared after alizarin red staining. Western blotting results showed that type Ⅰ collagen protein expression was significantly higher in the experimental group than in the control group (P < 0.01). ELISA quantitative analysis showed that osteocalcin was significantly higher in the experimental group than in the control group (P < 0.01). These findings suggest that whole bone marrow adherence method for culture of bone marrow mesenchymal stem cells is simple and practical and the cultured cells exhibit morphological and biological characteristics of osteoblasts after osteogenic induction.