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Genes 2016, 7(11), 103; doi:10.3390/genes7110103

Magnetofection Enhances Lentiviral-Mediated Transduction of Airway Epithelial Cells through Extracellular and Cellular Barriers

1
Department of Medical and Surgical Sciences, University of Foggia, V. L. Pinto 1, 71122 Foggia, Italy
2
Brainlab AG, Kapellenstrasse 12, 85622 Feldkirchen, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Michael Barry
Received: 16 September 2016 / Revised: 4 November 2016 / Accepted: 15 November 2016 / Published: 23 November 2016
(This article belongs to the Special Issue Gene Therapy)
View Full-Text   |   Download PDF [1938 KB, uploaded 23 November 2016]   |  

Abstract

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

Castellani, S.; Orlando, C.; Carbone, A.; Di Gioia, S.; Conese, M. Magnetofection Enhances Lentiviral-Mediated Transduction of Airway Epithelial Cells through Extracellular and Cellular Barriers. Genes 2016, 7, 103.

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