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Regeneration of a well-differentiated human airway surface epithelium by spheroid and lentivirus vector-transduced airway cells.

Abstract : BACKGROUND: Following injury to the airway epithelium, rapid regeneration of a functional epithelium is necessary in order to restore the epithelial barrier integrity. In the perspective of airway gene/cell therapy, we analyzed the capacity of human airway epithelial cells cultured as three-dimensional (3-D) spheroid structures to be efficiently transduced on long term by a pseudotyped lentiviral vector. The capacity of the 3-D spheroid structures to repopulate a denuded tracheal basement membrane and regenerate a well-differentiated airway epithelium was also analyzed. METHODS: An HIV-1-derived VSV-G pseudotyped lentiviral vector encoding the enhanced green fluorescent protein (eGFP) was used. Airway epithelial cells were isolated from mature human fetal tracheas and airway xenografts, cultured as 3-D spheroid structures, and either transduced at multiplicity of infection (MOI) 10 and 100 or assayed in an ex vivo and in vivo model to evaluate their regeneration capacity. RESULTS: An in vivo repopulation assay in SCID-hu mice with transduced isolated fetal airway epithelial cells shows that lentiviral transduction does not alter the airway reconstitution. Transduction of the 3-D spheroid structures shows that 12% of cells were eGFP-positive for up to 80 days. In ex vivo and in vivo assays (NUDE-hu mice), the 3-D spheroid structures are able to repopulate denuded basement membrane and reconstitute a well-differentiated human airway surface epithelium. CONCLUSIONS: The efficient and long-term lentiviral transduction of 3-D spheroid structures together with their capacity to regenerate a well-differentiated mucociliary epithelium demonstrate the potential relevance of these 3-D structures in human airway gene/cell therapy.
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https://www.hal.inserm.fr/inserm-00145977
Contributor : Philippe Birembaut <>
Submitted on : Monday, May 14, 2007 - 9:15:47 AM
Last modification on : Wednesday, September 16, 2020 - 4:54:13 PM

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Nicolas Castillon, Aurélie Avril-Delplanque, Christelle Coraux, Christophe Delenda, Bruno Péault, et al.. Regeneration of a well-differentiated human airway surface epithelium by spheroid and lentivirus vector-transduced airway cells.. The Journal of Gene Medicine, Wiley, 2004, 6 (8), pp.846-56. ⟨10.1002/jgm.570⟩. ⟨inserm-00145977⟩

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