Transfection of rat bone marrow stromal cells with human bone morphogenetic protein-7 recombinant adenovirus and its expression / 中国地方病学杂志

ABSTRACT

Objective To investigate the expression of human bone morphogenetic protein-7(hBMP-7)in rat bone malTOW stromal cells(BMSC)with a recombinant adenovirai vector carrying the hBMP-7 gene(Ad-hBMP-7) and study the effecta of Ad-hBMP-7 transfection on BMSC difierentiation.in order to explore the possibility for hBMP-7 gene therpy.Methods The rat BMSC cultured in vitro.They were divided into 3 groupsuntreated group,Ad-hBMP-7 and Ad-GFP transduced treated group.The rat BMSC were transfected by Ad-hBMP-7 and Ad-GFP. The expression of hBMP-7 was detected by RT-PCR and Western blot analysis.and the alkaline phosphatage (ALP)activity of the BMSC was observed.ResulIs In the Ad-hBMP-7 transduced treated group.hBMP-7 mRNA expression wag manifested detected by RT-PCR(470 bp),Westem blot analysis demonstrated that these cells indeed produced the hBMP-7 protein(Mr.15×103);10 days after transduction treatment,most of the BMSC were had brown black particles stained positively by ALP activity.But in Ad-GFP transduced treated group and untreated group they did not.Conclusions Ad-hBMP-7 could efficiently transfect BMSC and promote the conversion to osteoblast.The expression of hBMP-7 in rat BMSC provides a basis for hBMP-7 gene therapv.