基因电转染到三维重建的人皮肤组织

Title:Gene Electrotransfer in 3D Reconstructed Human Dermal Tissue

Volume: 16 Issue: 2

Author(s): Moinecha Madi, Marie-Pierre Rols, Laure Gibot

Affiliation:

关键词: 传递，质粒DNA，电穿孔，细胞外基质，基因治疗，皮肤，组织工程学

摘要: 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.

Cite this article as:

Moinecha Madi, Marie-Pierre Rols, Laure Gibot , Gene Electrotransfer in 3D Reconstructed Human Dermal Tissue, Current Gene Therapy 2016; 16 (2) . https://dx.doi.org/10.2174/1566523216666160331125052