骨质疏松大鼠骨髓基质细胞膜片制备过程中纤维连接蛋白表达检测  

Expression of Fibronectin in Osteoporotic Rat Bone Marrow Stromal Cell Sheet Constructed in vitro

作　　者：陈玉玲[1,2] 何梦娇 许雄程[2] 伍晓红 李艳芬[2,3] 陈超[2,3] 骆凯[2,3] CHEN Yuling;HE Mengjiao;XU Xiongcheng;WU Xiaohong;LI Yanfen;CHEN Chao;LUO Kai(Department of Orthodontics,The Hospital of Stomatology,Fujian Medical University,Fuzhou 350002,China;Key Laboratory of Stomatology of Fujian Provincial University,Fujian Medical University,Fuzhou 350005,China;Department of Periodontologhy,The Hospital of Stomatology,Fujian Medical University,Fuzhou 350002,China)

机构地区：[1]福建医科大学附属口腔医院正畸科,福州350002 [2]福建医科大学高校口腔医学重点实验室,福州350005 [3]福建医科大学附属口腔医院牙周科,福州350002

出　　处：《福建医科大学学报》2018年第6期374-378,共5页Journal of Fujian Medical University

基　　金：国家自然科学基金(81100761);福建省科技创新联合基金(2016Y9023);福建省教育厅科技项目(JA15199)

摘　　要：目的探讨骨质疏松大鼠骨髓基质细胞(BMSC)膜片体外构建过程中纤维连接蛋白(FN)的表达情况。方法体外分离培养骨质疏松大鼠BMSC,采用含50μg/mL维生素C的培养基连续培养构建细胞膜片,通过倒置显微镜和组织学染色对细胞膜片进行形态学观察;利用免疫荧光染色、实时荧光定量聚合酶链反应及Western-blot检测细胞外基质中FN在基因和蛋白水平的表达情况。结果采用含维生素C的培养基连续培养可获得含多层细胞及丰富胞外基质的BMSC细胞膜片。免疫荧光染色、实时荧光定量聚合酶链反应及Western-blot检测结果显示所构建的细胞膜片高表达FN。结论以浓度为50μg/mL的维生素C培养基连续培养可成功构建骨质疏松大鼠BMSC细胞膜片,膜片富含FN,有望应用于骨质疏松状态下牙槽骨缺损的治疗。Objective To detect the expression of fibronectin(FN)in osteoporotic rat bone marrow stromal cell(BMSC)cell sheet constructed in vitro.Methods BMSC were isolated and cultured from osteoporotic rats.Using culture medium supplemented with vitamin C,and osteoporotic rat BMSC cell sheet was tried to construct.Visual observation and histological staining were employed for morphological observation of the cell sheet.The mRNA and protein expression of FN in cell sheet were detected by Real-time PCR,Western-blot and immunofluorescence.Results Culturing in growth medium supplemented with vitamin C for about 10 days,osteoporotic rat BMSC cell sheet can be achieved.This white membranous structure was formed by layers of cells and abundant extracellular matrix.Real-time PCR,Western-blot and immunofluorescence result showed that FN was expressed at a high level in osteoporotic rat BMSC cell sheet.Conclusion Osteoporotic rat BMSC cell sheet can be constructed using vitamin C culture medium.The cell sheet is full of extracellular matrix and may be benefited for alveolar bone regeneration in osteoporosis.

关 键 词：骨质疏松 骨髓 骨髓细胞 细胞外基质 纤连蛋白类 

分 类 号：R332[医药卫生—人体生理学] R329.24[医药卫生—基础医学]