重组hBMP7基因转染成纤维细胞及其表达  

AdCMV-BMP7 transfected dermal fibroblasts express bone Morphogenic protein 7

作　　者：陈付国[1] 祝联[1] 刘德莉[1] 魏娴[1] 刘天一[1] 王敏[1] 柴岗[1] 王晓云[1] 周广东[1] 夏万尧[1] 崔磊[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海第二医科大学第九人民医院整复外科上海市组织工程研究重点实验室,上海200011

出　　处：《上海第二医科大学学报》2004年第4期238-241,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家重点基础研究发展规划资助项目("973"项目)(C19990543);上海市"重中之重"重点学科基金资助.

摘　　要：目的 探讨重组骨形态发生蛋白-7腺病毒载体(AdCMV-BMP7)在人真皮成纤维细胞获得稳定表达的可行性。方法重组腺病毒载体AdCMV-BMP7经293细胞包装、扩增后,转染人真皮成纤维细胞。转染48 h后以免疫组织化学、RT-PCR等方法检测hBMP7基因在人真皮成纤维细胞内的表达情况。结果 在合适的滴度下,重组腺病毒载体AdCMV-BMP7对成纤维细胞的增殖能力无明显影响。转染48 h后免疫细胞化学显示已转染AdCMV-BMP7的成纤维细胞内有大量的hBMP7表达,RT-PCR也检测到特异的hBMP7基因片段的表达。结论 重组腺病毒载体AdCMV-BMP7能够被转导入成纤维细胞内,并稳定表达hBMP7。Objective To explore the possibility of stable expression in transfected human dermal fibroblasts mediated by AdCMV-BMP7. Methods AdCMV-BMP7 was constructed via integration of hBMP7 cDNA into adenovirus vector. AdCMV-BMP7 was used to infect human dermal fibroblasts after encapsulation and proliferation in 293 cells. After 48 hours culture in vitro, expression of hBMP7 was examined with immunocytochemistry and RT-PCR. Results Massive hBMP expression was detected in cytoplasm of AdCMV-BMP7 infected fibroblasts with anti-hBMP7 monoclonal antibody. RT-PCR showed hBMP7 gene expression as early as 48 hours after AdCMV-BMP7 transfection. Conclusion hBMP7 gene can be transfected into dermal fibroblasts and be stably expressed.

关 键 词：骨形态发生蛋白-7 腺病毒 转基因技术 成纤维细胞 组织工程 RT-PCR 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]