大鼠骨髓间充质干细胞成骨诱导分化的超微结构变化

Ultrastructural changes following osteogenic differentiation of rat bone marrow mesenchymal stem cells

背景:目前对骨髓间充质干细胞成骨诱导分化的超微结构观察的报道甚少。目的:采用全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,成骨诱导并染色鉴定,利用电镜观察诱导前后细胞超微结构变化特点。方法:全骨髓贴壁法体外分离、培养、纯化大鼠骨髓间充质干细胞,流式细胞仪检测细胞表面标记物,成骨染色鉴定向成骨方向诱导分化,扫描电镜及透射电镜观察成骨诱导前后细胞超微结构变化。结果与结论:培养的第3代骨髓间充质干细胞纯度高、活力强,成骨诱导后的碱性磷酸酶活性染色、钙化结节染色均呈阳性扫描电镜及透射电镜观察显示,经向成骨细胞诱导分化后,细胞形态铺展,不规则,其线粒体、粗面内质网、空泡明显增多表明细胞功能活跃。

BACKGROUND： There are few studies on ultrastructure characteristics of osteoblasts after differentiation of bone marrow mesenchymal stem cells （BMSCs） OBJECTIVE： By using the whole bone marrow adherence method, to isolate, culture, purified BMSCs from rats, and to observe the ultrastructural changes after osteogenic differentiation of rat BMSCs. METHODS： BMSCs from rats were isolated, cultured and purified by the whole bone marrow adherence method; for morphology observations, cell surface markers were assessed by flow cytometry, and BMSCs were induced to differentiate into osteoblasts and stained to identification. Scanning electron microscope and transmission electron microscope were used to observe the ultrastructural changes of cells before and after induction. RESULTS AND CONCLUSION： The third generation cultured BMSCs had high purity and strong vitality. After induction, BMSCs were positive in bone alkaline phosphatase activity and calcified nodules staining. Scanning electron microscope and transmission electron microscope observation showed that after osteogenic induction, the cells spread, arranged irregularly, and were rich of organelles; and the cell mitochondria, rough endoplasmic reticulum, vacuoles increased significantly, indicating that the cells were functionally active.