Gene electrotransfer into the skin is of particular interest for the development
of medical applications including DNA vaccination, cancer treatment, wound
healing or treatment of local skin disorders. However, such clinical applications are
currently limited due to poor understanding of the mechanisms governing DNA electrotransfer
within human tissue. Nowadays, most studies are carried out in rodent models
but rodent skin varies from human skin in terms of cell composition and architecture.
We used a tissue-engineering approach to study gene electrotransfer mechanisms
in a human tissue context. Primary human dermal fibroblasts were cultured according to the self-assembly method to produce
3D reconstructed human dermal tissue. In this study, we showed that cells of the reconstructed cutaneous tissue were
efficiently electropermeabilized by applying millisecond electric pulses, without affecting their viability. A reporter gene
was successfully electrotransferred into this human tissue and gene expression was detected for up to 48h. Interestingly,
the transfected cells were solely located on the upper surface of the tissue, where they were in close contact with plasmid
DNA solution. Furthermore, we report evidences that electrotransfection success depends on plasmid mobility within tissue-
rich in collagens, but not on cell proliferation status. In conclusion, in addition to proposing a reliable alternative to
animal experiments, tissue engineering produces valid biological tool for the in vitro study of gene electrotransfer mechanisms
in human tissue.
Keywords: Delivery, Plasmid DNA, Electroporation, Extracellular matrix, Gene therapy, Skin, Tissue engineering.
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