大鼠骨髓间充质干细胞膜片的构建及其特性  

Construction of rat bone marrow mesenchymal stem cell sheet and study on it's biological properties

作　　者：安玉林[1] 那思家[1] 韦蔚 陈哲[1] 王国轩[1] 杜建宇[1] 金岩[1] 

机构地区：[1]第四军医大学口腔医学院组织工程中心,西安710032 [2]解放军359医院

出　　处：《实用口腔医学杂志》2012年第3期273-278,共6页Journal of Practical Stomatology

基　　金：国家高技术研究发展计划(863项目)(编号:2006AA02A119)

摘　　要：目的:探讨大鼠骨髓间充质干细胞膜片的构建和基本生物学特性。方法:从GFP-SD荧光大鼠体内提取原代骨髓并培养骨髓间充质干细胞(BMMSCs),鉴定后用于体外构建BMMSCs细胞膜片,用荧光显微镜、倒置显微镜和HE染色对膜片进行形态学检测,并提取膜片RNA用于RT-PCR实验,检测相关促愈合因子的表达情况。结果:BMMSCs成功分离、培养和鉴定。体外诱导培养2周后获得白色膜状细胞膜片,并可用机械力刮下。显微观察显示细胞呈长梭形重叠生长,染色显示细胞间紧密连接并分泌大量细胞外基质(ECM)。RT-PCR结果显示,膜片细胞与正常培养的BMMSCs相比,高表达TGF-β、FGF、VEGF和ColⅠ等细胞因子和蛋白,但在诱导1周和2周时的表达量无显著差异。结论:本方法能够较为稳定的在体外构建BMMSCs膜片,该膜片显著高表达TGF-β等促愈合因子。Objective： To construct bone marrow mesenchymal stem cell sheet and characterize it＇s fundamental biological property.Methods： Bone marrow mesenchymal stem cells（BMMSCs） were obtained from GFP-SD rats by operation and were used to construct BMMSC sheet.Fluorescence microscope,inverted microscope and HE staining were employed for morphological observation of the sheet.mRNA expression of TGF-β,FGF,VEGF and ColⅠ in the sheet was examined by RT-PCR.Results： After 2-week in vitro culture of BMMSCs a white membranaceous sheet was formed and was detachable by mechanical force.BMMSCs grew in multilayer and retained their fibroblastic spindle shape observed under microscope.HE staining showed abundant extracellular matrix（ECM） in the sheet.RT-PCR results indicated that the expression of the cytokines increased after 1 and 2 week culture.Conclusion： BMMSC sheet can be constructed by in vitro culture of BMMSCs.The sheet expresses high level of healing-promoting cytokines such as TGF-β and so on.

关 键 词：骨髓间充质干细胞 细胞膜片 细胞因子 愈合 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]