Title: Study on adenovirus-mediated gene transfer of basic fibroblast growth factor to rat's fetal smooth muscle cells in vitro
Abstract: Objective To investigate the efficiency and the effects of adenovirus-mediated gene transfer of basic fibroblast growth factor(bFGF)to rat's smooth muscle cells(SMCs)in vitro.Methods A replication-deficient recombinant adenovirus vector coding for bFGF(Ad.bFGF)was constructed and transferred into SMCs cultured in vitro,which were isolated and acquired from fetal rat's stomach.The efficiency of transfection was estimated by X-gal staining.On 1,2,3,5,7d after infection,the effect of Ad.bFGF on SMCs proliferation was evaluated by MTT assay.Also,bFGF protein expression was detected in the cell culture medium by ELISA on 1,2,3,5,8d after transfection.Results The constructed Ad.bFGF was a very efficient vector.When MOI reached 100,the ratio of infected cells arrived at 95% high in vitro.MTT assay revealed a significant increase in cell number of SMCs transfected with Ad.bFGF as compared with the cells transfected with Ad.LacZ and untransfected(PBS).After gene transfection,bFGF protein expressed in SMCs and secreted into the cell culture medium,which could be detected by ELISA.Conclusion A high efficient recombinant adenovirus vector coding for bFGF is successfully constructed.After transferred into SMCs in vitro,bFGF could be detected expressing and secreting in culture medium.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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