中国组织工程研究与临床康复2011，Vol.15Issue(23)：4303-4306,4.DOI:10.3969/j.issn.1673-8225.2011.23.028
携带肾上腺髓质素基因腺病毒体外转染骨髓间充质干细胞及其蛋白的表达
Adenovirus carrying adrenomedullin gene for transfection of bone marrow mesenchymal stem cells in vitro and their protein expression
摘要
Abstract
BACKGROUND: Adrenomedullin is one of potential growth factor that can promote cellular differentiation, growth and reduce apoptosis.OBJECTIVE: To investigate the possibility of bone marrow mesenchymal stem cells transfected by adenovirus vector carrying adrenomedullin (Ad-ADM), and to determine the protein expression of adrenomedullin following transfection. METHODS: The 293A cells were cultivated and the Ad-ADM was amplified. The experiment was divided into three groups: control, blank vector and Ad-ADM transfection. Cells in three groups were cultured with serum-free L-DMEM, Ad-lacZ and Ad-ADM, respectively after they reached 50％-60％ confluence. The virus titer of amplified Ad-ADM was detected by plaque methods. Bone marrow mesenchymal stem cells were isolated and expanded using the preplating methods. The efficiency of Ad-GFP transfected mesenchymal stem cells in vitro was determined. Adrenomedullin mRNA expression was detected with reverse transcription-polymerase chain reaction following transfection.RESULTS AND CONCLUSION: The virus titer of amplified Ad-ADM was 1.1×109 pfu/mL by plaque methods. After bone marrow mesenchymal stem cells were infected with Ad-ADM, the expression of adrenomedullin mRNA was significantly increased. The expression of adrenomedullin mRNA was not observed in the control group and empty vector group. Quantitative results showed that the expression gradually increased from day 1, reached the peak at day 7, and was still detectable at day 11. The Ad-ADM can successfully transfect bone marrow mesenchymal stem cells. Following transfection, bone marrow mesenchymal stem cells highly express adrenomedullin.关键词
肾上腺髓质素/293细胞/噬斑试验/骨髓间充质干细胞/腺病毒载体分类
医药卫生引用本文复制引用
林茂海,苏津自,蔡文钦,郑礼裕,许昌声,王华军..携带肾上腺髓质素基因腺病毒体外转染骨髓间充质干细胞及其蛋白的表达[J].中国组织工程研究与临床康复,2011,15(23):4303-4306,4.基金项目
福建医科大学附属第一医院"院学科带头人"科研基金(JXK200704). （JXK200704）
