Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β_1 (TGF-β_ 1) cDNA, and the expression of the encoded prote...Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β_1 (TGF-β_ 1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerat ive lumbar disc disease. The cells were cultured and directly infected by two a denoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence pro tein (EGFP) gene (marker gene) and Ad/CMV-TGF-β_1 containing the potentially therapeutic TGF-β_1 gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-β_1). Results Culture cells transducted by Ad/CMV-EGFP showed specific green fluorescence und er the fluoroscope and expression sustained for at least 4 weeks. When infe cted by Ad/CMV-TGF-β_1, approximally 30% of cultured cells were staind brown (+) with TGF-β_1 staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene , TGF-β_1, by using an adenoviral vector to human degenerative lumbar interve rtebral disc cells.展开更多
文摘Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β_1 (TGF-β_ 1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerat ive lumbar disc disease. The cells were cultured and directly infected by two a denoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence pro tein (EGFP) gene (marker gene) and Ad/CMV-TGF-β_1 containing the potentially therapeutic TGF-β_1 gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-β_1). Results Culture cells transducted by Ad/CMV-EGFP showed specific green fluorescence und er the fluoroscope and expression sustained for at least 4 weeks. When infe cted by Ad/CMV-TGF-β_1, approximally 30% of cultured cells were staind brown (+) with TGF-β_1 staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene , TGF-β_1, by using an adenoviral vector to human degenerative lumbar interve rtebral disc cells.
