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Allo, JC ; Midoux, P ; Merten, M ; Souil, E ; Lipecka, J ; Figarella, C ; Monsigny, M ; Briand, P ; Fajac, I

Efficient gene transfer into human normal and cystic fibrosis tracheal gland serous cells with synthetic vectors

American Journal of Respiratory Cell and Molecular Biology 22 (2) 166-175

par Administrateur - publié le , mis à jour le

Abstract :

Submucosal gland serous cells are believed to play a major role in the physiopathology of cystic fibrosis (CF) and may represent an important target for CF gene therapy. We have studied the efficiency of re porter gene transfer into immortalized normal (MM-39) and CF (CF-KM I) human airway epithelial gland serous cells using various synthetic vectors : glycosylated polylysines (glycofectins), polyethylenimine (PEI) (25 and 800 kD), lipofectin, and lipofectAMINE. In both cell lines, a high luciferase activity was achieved with various glycofectins, with PEI 25 kD, and with lipofectAMINE. After three transfections applied daily using a-glycosylated polylysine, 20% of the cells were transfected. At 24 h after CF trans membrane conductance regulator (CFTR) gene transfer into CF-KM4 cells using a-glycosylated poly lysine, the immunolocalization of CFTR was analyzed by laser scanning confocal microscopy and the transgenic CFTR was detected by an intense labeling of the plasma membrane.