›› 2013, Vol. 4 ›› Issue (4): 169-175.

    Next Articles

Expression of hOPG in PDLCs of dog infected by recombinant adenoviral vector Ad5-hOPG-EGFP

  

  • Received:2013-10-14 Revised:2013-12-01 Online:2013-12-25 Published:2014-01-09

Abstract: Objective:To evaluate the expression of human osteoprotegerin (hOPG) in periodontal ligament cells (PDLCs) of Beagle dog in order to prepare seed cells with high expression of hOPG gene for periodontal tissue regeneration experiment in later stage. Methods:PDLCs of Beagle dogs were isolated and cultured. Then, adenovirus vector carrying hOPG gene was structured in vitro and transfected into canine PDLCs. Expression of hOPG was observed by flow cytometry, real-time RT-PCR, ELISA and Western Blot. Results:Recombinant adenovirus Ad5-hOPG-EGFP was successfully transfected PDLCs of Beagle dogs. More than 80 per cent of transfected cells positively expressing the green fluorescene protein could be observed under fluorescence microscope and detected by flow cytometry. Effective expression of hOPG was also observed by real-time RT-PCR, ELISA and Western Blot, which proved the successful transfect hOPG into canine PDLCs. Conclusions:hOPG was successfully expressed in PDLCs of Beagle dog which were infected by recombinant adenoviral vector Ad5-hOPG-EGFP.