中国组织工程研究2012，Vol.16Issue(23)：4273-4278,6.DOI:10.3969/j.issn.1673-8225.2012.23.019
人骨形态发生蛋白2和人成纤维细胞生长因子2双基因共表达腺病毒载体转染人骨髓基质干细胞后的细胞增殖
In vitro proliferation of human bone marrow stromal cells after transfection by denovirus vector co-expressing human bone morphogenetic protein-2 and human fibroblast growth factor 2 genes
摘要
Abstract
BACKGROUND: Transfection of exogenous gene into tissue-engineered seed cells via recombinant adenovirus vector is the key to gene therapy of bone defects.OBJECTIVE: To investigate the effect of gene transfection on the proliferation of human bone marrow stromal stem cellsthat tranfected with adenovirus vectors co-expressing human bone morphogenetic protein-2 (hBMP-2) and humanfibroblast growth factor 2 (hFGF-2).METHODS: The Ad-hBMP2-IRES-hFGF2 plasmids were transfected into human bone marrow stromal stem cells. Theefficiency of transfection was evaluated by fluorescence microscope. Reverse transcriptase polymerase chain reactionwas used to observe the successful transcription of hBMP-2 and hFGF-2 cNDA in bone marrow mesenchymal stem cells.Western blot assay was used to identify the protein expression of hBMP-2 and hFGF-2 genes. The cellular viability wasdetermined by trypan blue staining and the changes of the cell proliferation were observed by flow cytometry assay.RESULTS AND CONCLUSION: The expression of hBMP-2 and hFGF-2 genes was confirmed at mRNA and proteinlevels. The cell viability was not changed after transfection. The flow cytometry assay showed that the percentage ofproliferating cells was increased significantly in cell cycle. The Ad-hBMP2-IRES-hFGF2 plasmids could transfect humanbone marrow stromal stem cells with high transfection efficiency and promote cell proliferation.关键词
腺病毒/人骨形态发生蛋白2/人成纤维细胞生长因子2/骨髓基质干细胞/基因转染/干细胞分类
医药卫生引用本文复制引用
郭伟韬,王辉,刘思景,曾荣,肖启贤,陈子秋,黄云,王斌,胡资兵..人骨形态发生蛋白2和人成纤维细胞生长因子2双基因共表达腺病毒载体转染人骨髓基质干细胞后的细胞增殖[J].中国组织工程研究,2012,16(23):4273-4278,6.基金项目
广东省自然科学基金资助项目(S2011010004096) （S2011010004096）
广东省医学科研基金资助项目(A2009477,A2010431). （A2009477,A2010431）
