中国组织工程研究Issue(45)：7847-7854,8.DOI:10.3969/j.issn.2095-4344.2013.45.005
骨髓间充质干细胞中软骨调节素Ⅰ的稳定上调表达
Stably upregulating expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND:Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰmaybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately. <br> OBJECTIVE:To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells. <br> METHODS:Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in celllines. <br> RESULTS AND CONCLUSION:The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successful y constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells.关键词
干细胞/骨髓干细胞/软骨调节素Ⅰ/骨髓间充质干细胞/软骨细胞/载体/生物因子/组织工程/国家自然科学基金/干细胞图片文章分类
医药卫生引用本文复制引用
周连仲,崔成华,冯亚,邢双春,翟立杰..骨髓间充质干细胞中软骨调节素Ⅰ的稳定上调表达[J].中国组织工程研究,2013,(45):7847-7854,8.基金项目
国家自然科学基金面上项目(812717202012)；辽宁省教育厅科学研究一般项目(L20123262012)@@@@the National Natural Science Foundation of China, No.812717202012 （812717202012）
the General Program of Science Research of Liaoning Educational Bureau, No. L20123262012 （）
