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

    Marcel Huber, Daniel Hohl, Alain Limat, Ernst Wagner
    The study investigated three nonviral transfection systems for gene transfer in human keratinocytes, with a focus on the adenovirus-enhanced receptor-mediated (AVET) system. The AVET system showed superior efficiency, achieving a transfection rate of 28.8%, compared to lower rates with SuperFect and no success with PrimeFector. The presence of adenovirus was crucial for effective gene delivery. The study also found that slowly adhering keratinocytes were transfected more effectively, suggesting stem cells were less efficiently transfected. The AVET system was able to correct the enzymatic defect in keratinocytes from lamellar ichthyosis patients, but was unsuccessful in organotypic cultures due to barriers like the cornified layer. Overall, the AVET system was a powerful tool for transient in vitro transfection of keratinocytes, with potential applications in gene therapy for skin diseases.
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