LIM矿化蛋白1慢病毒载体转染后大鼠骨髓基质干细胞的分化  

Differentiation of bone mesenchymal stem cells of rat following lentivirus transfection with LIM mineralization protein-1

作　　者：张姝江[1] 谭丽[2] 陈艺[1] 白波[1] 

机构地区：[1]广州医科大学附属第一医院广东省骨科矫形技术与植入材料重点实验室,510120 [2]广州医科大学附属第一医院血液科,510120

出　　处：《中华关节外科杂志（电子版）》2015年第5期623-626,共4页Chinese Journal of Joint Surgery(Electronic Edition)

基　　金：国家自然科学基金(81371978);广东省自然科学基金(S2013010014787)

摘　　要：目的探讨慢病毒转染大鼠骨髓基质干细胞(BMSCs)过表达LIM矿化蛋白1(LMP-1)对BMSCs分化能力的影响。方法构建LMP-1基因慢病毒载体,转染原代培养的大鼠MSCs,荧光显微镜下观察转染效果,MTT法检测转染后细胞增殖,常规方法诱导过表达LMP-1的BMSCs成骨和成软骨分化,以免疫组织化学染色观察成骨诱导后细胞分泌骨钙素的情况以及用阿利新蓝染色鉴定成软骨诱导后细胞分泌氨基多糖的情况。结果感染复数(MOI)值为12时即可获得良好的转染效果,细胞转染后增殖正常,成骨诱导后,骨钙素染色阳性;成软骨诱导后,阿利新蓝染色阳性。结论lv-lmp-1可以有效转染大鼠BMSCs,转染后细胞的多向分化能力没有受到影响。Objective To explore the influences of LIM mineralization protein-1(LMP-1) overepression by lentivirus on the differentiation ability of bone mesenchymal stem cells ( BMSCs) of rat. Methods The vector for LMP-1 gene was constructed by lentivirus, which transfected the BMSCs in vitro. The transfected cells were observed by fluorescence microscope, and MTT assay was used to determine cell proliferation.The LMP-1-overexpressed BMSCs were induced by regular protocols to differentiate into osteoblasts and chondrocytes. Immunochemistry staining was performed to determine osteocalcin expression, while alcian blue staining was performed to determine aggrecan expression.Results The cells were well transfected at multiplicity of infection ( MOI ) value of 12.MTT assay showed a normal proliferation curve of transfected BMSCs. After osteogenic and chondrogenic inductions, osteocalcin staining and alcian blue staining were positive respectively.Conclusion Lv-lmp-1 vector can effectively transfect BMSCs of rat which would then overexpress LMP-1, and the muti-differentiation ability of BMSCs is not influenced.

关 键 词：慢病毒属 转染 LIM矿化蛋白1 细胞分化 骨髓 间充质干细胞 

分 类 号：R68[医药卫生—骨科学]