Efficient In Vitro Transfection of Human Keratinocytes with an Adenovirus-Enhanced Receptor-Mediated System

The study investigated the efficiency of three nonviral transfection systems for gene delivery in human keratinocytes, with a focus on the adenovirus-enhanced transferrin-mediated (AVET) system. The AVET system demonstrated a transfection efficiency of 28.8%, outperforming SuperFect and PrimeFector. The presence of adenovirus was crucial for effective gene delivery. The system was more effective in slowly adhering keratinocytes, suggesting stem cells were less efficiently transfected. The AVET system corrected the enzymatic defect in keratinocytes from lamellar ichthyosis patients, achieving enzyme activity levels similar to normal keratinocytes. However, it was unsuccessful in organotypic cultures from outer root sheath cells, likely due to the cornified layer or receptor inaccessibility. Overall, the AVET system was a powerful tool for transient in vitro transfection of keratinocytes, with potential applications in gene therapy for skin diseases.