Huang Qiusheng, Lu Hanqiang, Zhou Yuepeng, He Qinghua, Sun Xianglan, Jiang Ping, Zhang Zhijian
Department of Otolaryngology, the Affiliated Hospital of Jiangsu University, Zhenjiang, 212002, China.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2012 Jun;26(11):490-3, 498.
To establish an in vitro method to culture mesenchymal stem cells(MSCs) derived from human nasal mucosa, and explore their stemness and differentiation potential.
Based on the observation of distribution of MSCs in human nasal mucosa, we cultured and proliferated MSCs in vitro and identified the expression of stem cell markers on them including Nestin, CD133, Vimentin and Sa114 with immunofluorescence. The MSCs were induced to differentiate to osteoblasts with medium containing dexamethasone, ascorbic acid and beta sodium glycerophosphate, and to neurons with Neurobasal medium containing B27, ATRA and TSA. Histochemistry and immunofluorescence were applied to evaluate the differentiation.
The nestin and vimentin immunofluorescence-positive MSCs existed extensively in human nasal mucosa. While the MSCs were cultured in the osteogenic-inducing medium, activities of alkaline phosphatase were increased significantly, and bone nodules were found on the surface of the osteoblasts by alizarin red staining. After the induction by neural-inducing medium, the MSCs adopted neuron like appearance with many slim protrusions interconnected as a network. The induced cells expressed neural markers NF-200 and BM88 strongly.
The MSCs derived from human nasal mucosa are multipotent stem cells and can be utilized as seed cells to repair bone or neural injury.
建立一种体外培养人鼻黏膜间充质干细胞（MSCs）的方法，并探究其干性和分化潜能。
基于对人鼻黏膜中MSCs分布的观察，我们在体外培养并扩增MSCs，并用免疫荧光法鉴定其干细胞标志物Nestin、CD133、波形蛋白和Sca-1的表达。将MSCs用含地塞米松、抗坏血酸和β-甘油磷酸钠的培养基诱导分化为成骨细胞，并用含B27、全反式维甲酸（ATRA）和曲古抑菌素A（TSA）的神经基础培养基诱导分化为神经元。应用组织化学和免疫荧光法评估分化情况。
Nestin和波形蛋白免疫荧光阳性的MSCs广泛存在于人鼻黏膜中。当MSCs在成骨诱导培养基中培养时，碱性磷酸酶活性显著增加，茜素红染色显示成骨细胞表面有骨结节形成。经神经诱导培养基诱导后，MSCs呈现出神经元样外观，有许多细长的突起相互连接成网络。诱导后的细胞强烈表达神经标志物NF-200和BM88。
人鼻黏膜来源的MSCs是多能干细胞，可作为种子细胞用于修复骨或神经损伤。
