稳定表达人骨形态发生蛋白2基因骨组织工程种子细胞的构建.docVIP

稳定表达人骨形态发生蛋白2基因骨组织工程种子细胞的构建 俞莉敏1，马俊轩2，李继云1，于滨生1(1北京大学深圳医院脊柱外科，广东省深圳市 518036；2深圳市脊柱外科重点实验室，广东省深圳市 518036) 引用本文：俞莉敏，马俊轩，李继云，于滨生. 稳定表达人骨形态发生蛋白2基因骨组织工程种子细胞的构建[J].中国组织工程研究，2017，21():2722-2728. DOI: 10.3969/j.issn.2095-4344.2017.17.016 ORCID: 0000-0002-6486-8656(俞莉敏) 文章快速阅读： 文题释义： 基因转染间充质干细胞：通过将成骨基因导入细胞，不同于重组细胞因子的爆发性释放，可实现高活性成骨蛋白的持久稳定表达及生理性释放。其中骨髓来源间充质干细胞在成骨诱导下可分化为成骨细胞，介导骨组织的再生，将骨形态蛋白基因导入间充质干细胞，不仅可实现骨形态蛋白的持续性缓释，也提供了骨组织修复所需要的有成骨分化潜能的干细胞，采用这类稳定表达骨形态蛋白基因的干细胞促进骨组织修复，已在动物实验得到了证实。 稳定表达人骨形态发生蛋白2基因的骨组织工程种子细胞构建：Construction of seed cells with the stable expression of human bone morphogenetic protein 2 gene for bone tissue engineering Yu Li-min1, Ma Jun-xuan2, Li Ji-yun1, Yu Bin-sheng1 (1Department of Spine Surgery, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong Province, China; 2Shenzhen Key Laboratory of Spine Surgery, Shenzhen 518036, Guangdong Province, China) Abstract BACKGROUND: Because of the non-homology of protein and gene between human and animals, to promote osteogenesis or spinal fusion of animals by construction of tissue-engineered bone with the human gene has influenced the experimental validation. OBJECTIVE: To construct the seed cell line for bone tissue engineering with stable expression of human bone morphogenetic protein 2 (hBMP2). METHODS: The full-length hBMP2 gene was cloned from human muscle tissues by nested RT-PCR and transfected to human bone marrow mesenchymal stem cells (hBMSCs) with lipidosome. The transfected hBMSCs were cultured with G418 in vitro to screen and purify the cells. A series of analyses such as RT-PCR, dot-ELISA, immunohistochemstry and alkaline phosphatase activity analysis were performed to evaluate the situation of hBMP2 expression and secretion at 48 hours and 3 weeks after the transduction. hBMSCs transduced with empty plasmid and the normal hBMSCs served as positive control and blank control groups, respectively, which were used for observation of cell growth, proliferation and biological characterist