鼻黏膜外胚层间充质干细胞诱导分化成骨细胞实验分析.docVIP

鼻黏膜外胚层间充质干细胞诱导分化成骨细胞实验分析 　　[摘要] 目的 探讨鼻黏膜外胚层间充质干细胞经诱导分化为成骨细胞的效果。 方法 收集SD大鼠鼻黏膜，进行鼻黏膜外胚层间充质干细胞体外培养。对细胞表面标志物CD45-FITC、CD14-PE、CD105-PE、CD34-PE、CD71-PE、CD34-PE、CD44-PE进行鉴定，并对细胞进行成骨诱导。成骨诱导分化后检测细胞在不同时间的碱性磷酸酶活性变化情况，并对细胞进行茜素红染色，观察细胞诱导分化效果。 结果 大鼠鼻黏膜外胚层间充质干细胞经成骨诱导分化后，碱性磷酸酶活性在诱导4 d后显著增加，并在7 d达到最大值，之后逐渐下降。成骨诱导至21 d，对大鼠鼻黏膜外胚层间充质干细胞进行茜素红染色之后可观察到结果呈现出阳性，细胞外基质中出现钙沉积以及钙结节形成。 结论 在大鼠鼻黏膜中存在一定的外胚层间充质干细胞，该细胞具有分化潜能，经诱导分化可分化为成骨细胞，具有一定的临床应用前景。 　　[关键词] 外胚层间充质干细胞；鼻黏膜；诱导；分化；成骨细胞 　　[中图分类号] R329.2 [文献标识码] A [文章编号] 1674-4721（2016）08（b）-0029-05 　　[Abstract] Objective To investigate the effect of the induction and differentiation of ectomesenchymal stem cells derived from nasal mucosa into osteoblast. Methods The nasal mucosa of SD rat was collected and the ectomesenchymal stem cells derived from nasal mucosa were cultured in vitro. The cell surface markers CD45-FITC， CD14-PE， CD105-PE， CD34-PE， CD71-PE， CD34-PE， CD44-PE were identified， and the cells were taken osteogenic induction. After induction and differentiation of osteoblast， the changes of alkaline phosphatase activities in different time were detected， and the cells were taken alizarin red staining， the effects of induction and differentiation of cells were observed. Results Through induction and differentiation of osteoblast， the alkaline phosphatase activities of ectomesenchymal stem cells derived from nasal mucosa were increased significantly after induction for 4 d， which reached maximum value at 7 d and decreased gradually later. After osteogenic induction for 21 d， the ectomesenchymal stem cells derived from nasal mucosa by alizarin red staining showed positive results， extracellular matrix showed calcium deposit and the formation of calcium nodule. Conclusion There are some ectomesenchymal stem cells′ in the nasal mucosa of rats， the cells have the ability of differentiation and can be differentiated into osteoblasts through induction and differentiation， which has certain clinical application prospects. 　　[Key words] Ectomese