Hogquist, Stephen Joseph. Utilizing a Franz cell device for the electroporation of adherent cells with nanoporous alumina substrates. Retrieved from https://doi.org/doi:10.7282/t3-dcrw-4b86
DescriptionElectroporation is the technique of applying an electric field to a cell to achieve temporary membrane permeabilization and allow the passage of drugs, chemicals, or DNA through the compromised cell membrane. Electroporation technology is presently an area of research interest due to its versatility and potential benefit to various investigational therapies including vaccines and cell and gene therapies. The transfection efficiency and cell viability following electroporation experiments depend strongly on various parameters such as the applied electric field, cell type used, and the contents and electrical characteristics of the buffer solution through which a field is applied. This study aims to demonstrate a technique by which a lower voltage can be used to electroporate cells with the goal of increasing cell viability and permeabilization efficiency with plasmid DNA and propidium iodide when compared to existing techniques. This is achieved utilizing 3T3 cells attached to a biocompatible, 60µm thick nanoporous alumina membrane substrate with 100nm pores. Computational modeling predicts that applying an electric field through this nanoporous substrate in a conductive buffer solution results in electric field amplification near nanopores to achieve electroporation of cells adherent to the substrate, while maintaining a lower field strength elsewhere in the remaining area surrounding a cell. As many existing electroporation experiments work with cell suspensions in cuvettes to achieve electroporation, this technique presents a method to achieve electroporation of adherent cells. Results suggest that delivery cargo of interest can be electrophoretically driven through these nanopores and enter a cell with an electroporation pulse under select electric field conditions. Here, a technique is described to further investigate the application of nanoporous substrates in electroporation experiments. Furthermore, the impact of electric field strength amplification through alumina nanopores is demonstrated by the successful permeabilization of cell membranes with delivery markers.