Lentiviral vector-mediated transfection of bone morphogenetic protein 2 gene into endothelial progenitor cells from rat bone marrow
10.3969/j.issn.2095-4344.2014.32.019
- VernacularTitle:慢病毒介导骨形态发生蛋白2基因转染大鼠骨髓内皮祖细胞
- Author:
Xiuru YIN
;
Ling PEI
;
Zuodi LIANG
- Publication Type:Journal Article
- Keywords:
endothelial cells;
bone morphogenetic proteins;
lentivirus;
transfection
- From:
Chinese Journal of Tissue Engineering Research
2014;(32):5197-5202
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. The key point is to choose the proper cell, gene and vector. OBJECTIVE:To investigate the effectiveness and feasibility of bone morphogenetic protein 2 (BMP2) gene transfected into endothelial progenitor cells (EPCs) from rat bone marrow for gene therapy. METHODS:The EPCs were isolated, cultured and identified from the bone marrow of Sprague-Dawley rats. Empty vector (LV-eGFP) or BMP2 gene (LV-eGFP-BMP2) was transferred into EPCs by the constructed lentiviral vector (LV). We examined the transfection efficiency by eGFP fluorescence, BMP2 secretion by ELISA, BMP2 expression by Western blot, and compared the capacities of migration, proliferation and anti-apoptosis after transfection in the three groups of normal EPCs, empty vector-EPCs, and BMP2-EPCs. RESULTS AND CONCLUSION:The transfection efficiency of lentiviral vector was 90%. BMP2 gene-transferred EPCs secreted and expressed more BMP2 proteins (P<0.01), and showed enhanced anti-apoptotic ability (P<0.05). The proliferation and migration capacity did not change obviously (P>0.05). After successful transfection with lentivirus-BMP2 gene, EPCs can secrete and express more BMP2 protein and show enhanced anti-apoptotic ability without obvious influence on the proliferation and migration capacity.
