摘要
To investigate the osteogenic potential of adipose tissue-derived stromal cells (ADSCs) exposed to adenovirus containing hBMP-2 cDNA(Adv-hBMP-2) and provide an optimal cell source for gene therapy.Methods ADSCs were obtained from patients who received orthopaedic surgery or liposuction.Human adipose tissue underwent proteolytic digestion to obtain ADSCs.After exposure to Adv-hBMP-2,immunorecipitation and Western blotting and ELISA were performed for confirming BMP gene transduction.The apoptosis were observed by flow cytometry.ADSCs -Adv-hBMP-2 cells were injected into the hind limb of nude mice.Radiographic and histological evalution were performed on schedule to comfirm the bone formation.Results ADSCs were successfully isolated from human adip0ose tissue.The transfection efficiency was examined by X-gal staining.Western blotting and ELISA confirmed successful and perisistent BMP-2 production by ADSCs-Adv-hBMP-2 cells.The flow cytometry showed there was some negative effect on apoptosis of ADSCs after exposed to adenovirus containing cDNA,but the rate of apoptosis is not so high that compromised the treatment.X-ray and HE sections from hind limb of nude mice subject to injection of Adv-hBMP-2 cells confirmed bone formation at 2nd week.Conclusion Adipose tissue contains abundant ADSCs that can be transfected with BMP-2 gene,with ectopic bone formation.ADSCs may be an ideal source of cells for gene therapy.2 refs.
To investigate the osteogenic potential of adipose tissue-derived stromal cells (ADSCs) exposed to adenovirus containing hBMP-2 cDNA(Adv-hBMP-2) and provide an optimal cell source for gene therapy.Methods ADSCs were obtained from patients who received orthopaedic surgery or liposuction.Human adipose tissue underwent proteolytic digestion to obtain ADSCs.After exposure to Adv-hBMP-2,immunorecipitation and Western blotting and ELISA were performed for confirming BMP gene transduction.The apoptosis were observed by flow cytometry.ADSCs -Adv-hBMP-2 cells were injected into the hind limb of nude mice.Radiographic and histological evalution were performed on schedule to comfirm the bone formation.Results ADSCs were successfully isolated from human adip0ose tissue.The transfection efficiency was examined by X-gal staining.Western blotting and ELISA confirmed successful and perisistent BMP-2 production by ADSCs-Adv-hBMP-2 cells.The flow cytometry showed there was some negative effect on apoptosis of ADSCs after exposed to adenovirus containing cDNA,but the rate of apoptosis is not so high that compromised the treatment.X-ray and HE sections from hind limb of nude mice subject to injection of Adv-hBMP-2 cells confirmed bone formation at 2nd week.Conclusion Adipose tissue contains abundant ADSCs that can be transfected with BMP-2 gene,with ectopic bone formation.ADSCs may be an ideal source of cells for gene therapy.2 refs.
出处
《外科研究与新技术》
2005年第3期183-184,共2页
Surgical Research and New Technique
