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目的应用骨膜提取物诱导骨髓间充质干细胞(MSCs)分化为类成骨细胞,为骨组织工程提供充足的种子细胞。方法切取小鼠颅骨骨膜提取物,诱导小鼠骨髓间充质干细胞。诱导分化后的细胞采用ALP染色、RT-PCR、蛋白印迹技术进行检测。结果形态学观察显示,分化后的MSCs细胞形态从长梭形变成三角形、多角形或立方形;ALP活性结果显示,分化后的MSCs细胞ALP活性远高于未分化的MSCs;蛋白印迹显示,分化后的MSCs表达成骨细胞相关的BMP-2蛋白;RT-PCR结果表明,分化后的MSCs表达成骨细胞相关的4个基因Type(I)collagen、Osteocalcin、ALP和BMP-2,而未分化的MSCs则几乎不表达。结论骨膜提取物可以诱导小鼠骨髓间充质干细胞分化为类成骨细胞。
Objective To apply periosteum extracts to induce bone marrow mesenchymal stem cells (MSCs) to differentiate into osteoblasts and provide sufficient seed cells for bone tissue engineering. Methods Mouse skull periosteum extract was excised to induce mouse bone marrow mesenchymal stem cells. The differentiated cells were detected by ALP staining, RT-PCR and Western blotting. Results Morphological observation showed that the morphology of the differentiated MSCs changed from long fusiform to triangular, polygonal or cubic shape. ALP activity showed that the ALP activity of differentiated MSCs was much higher than that of undifferentiated MSCs. Western blotting showed that, Differentiated MSCs expressed osteoblast-associated BMP-2 protein. RT-PCR results showed that the differentiated MSCs expressed osteoblast-related four genes Type (I) collagen, Osteocalcin, ALP and BMP-2, Differentiated MSCs are almost not expressed. Conclusion Periosteum extract can induce mouse bone marrow mesenchymal stem cells to differentiate into osteoblasts.