Gene transfer to airway epithelial cells is hampered by extracellular (mainly mucus) and\ncellular (tight junctions) barriers. Magnetofection has been used to increase retention time of lentiviral\nvectors (LV) on the cellular surface. In this study, magnetofection was investigated in airway epithelial\ncell models mimicking extracellular and cellular barriers. Bronchiolar epithelial cells (H441 line)\nwere evaluated for LV-mediated transduction after polarization onto filters and dexamethasone (dex)\ntreatment, which induced hemicyst formation, with or without magnetofection. Sputum from cystic\nfibrosis (CF) patients was overlaid onto cells, and LV-mediated transduction was evaluated in the\nabsence or presence of magnetofection. Magnetofection of unpolarized H441 cells increased the\ntransduction with 50 MOI (multiplicity of infection, i.e., transducing units/cell) up to the transduction\nobtained with 500 MOI in the absence of magnetofection. Magnetofection well-enhanced LV-mediated\ntransduction in mucus-layered cells by 20.3-fold. LV-mediated transduction efficiency decreased\nin dex-induced hemicysts in a time-dependent fashion. In dome-forming cells, zonula occludens-1\n(ZO-1) localization at the cell borders was increased by dex treatment. Under these experimental\nconditions, magnetofection significantly increased LV transduction by 5.3-fold. In conclusion,\nthese results show that magnetofection can enhance LV-mediated gene transfer into airway epithelial\ncells in the presence of extracellular (sputum) and cellular (tight junctions) barriers, representing\nCF-like conditions.
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