CAJ | 학술논문

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。
목적：탐토만병독전염대서골수기질간세포（BMSCs）과표체LIM광화단백1（LMP－1）대BMSCs분화능력적영향。방법구건LMP－1기인만병독재체，전염원대배양적대서MSCs，형광현미경하관찰전염효과，MTT법검측전염후세포증식，상규방법유도과표체LMP－1적BMSCs성골화성연골분화，이면역조직화학염색관찰성골유도후세포분비골개소적정황이급용아리신람염색감정성연골유도후세포분비안기다당적정황。결과감염복수（ MOI）치위12시즉가획득량호적전염효과，세포전염후증식정상，성골유도후，골개소염색양성；성연골유도후，아리신람염색양성。결론lv－lmp－1가이유효전염대서BMSCs，전염후세포적다향분화능력몰유수도영향。
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.