Adenovirus-mediated VEGF gene in human bone marrow stromal cells.docVIP

 PAGE \* MERGEFORMAT 13 Adenovirus-mediated VEGF gene in human bone marrow stromal cells Of: Chen Bin, Song Yanbin, Li Yuhua, Guo-xian Pei [Abstract] [Objective] in cultured human bone marrow stromal cells (hBMSc), the VEGF165 gene adenovirus vector cotransfected hBMSc, observed its expression. [Method] co-transfection experiments were divided into 3 groups: VEGF165 and eGFP transfected group, empty adenovirus vector transfected and untransfected group. take adenovirus Ad VEGF165 (MOI = 20 pfu / cell) infection hBMSe. eGFP fluorescence expression under the microscope to determine infection efficiency and cell growth, RT PCR detection ELISA test were used for observation and Westernblot gene expression. [results] MOI = 20 pfu transfection efficiency reached 80% or more, take this value transfection titer. extracted total RNA of good quality. to double-stranded cDNA as template amplified VEGF165 gene product by 1% agarose gel electrophoresis, can be seen 549 bp (GAPDH) 2 bands. VEGF165 were observed in the expression of transfected, empty vector group and control group did not detect VEGF165 expression. ELISA results showed that gene transfer group transfected with empty vector and untransfected group difference between the results was statistically significant (P lt;0.05). Western blot shown: transfected in 50 KD occurring at the characteristic bands, while empty vector group and control group was no band. [Conclusion] exogenous VEGF genes in human bone marrow stromal cells (hBMSc) expression within a stable, cultured under the experimental conditions of hBMSc typical morphological characteristics of osteoblasts and strong biological activity, differentiate into osteoblasts higher efficiency cells can meet the requirements of bone tissue engineering. [Keywords:] bone marrow stromal cells, VEGF, green fluorescent protein gene transfection Abstract: [Objective] To investigate the possibility of stable expression of VEGF ~ 65 in human bone marrow stroma