Gene electrotransfer into mammalian cells using commercial cell culture inserts with porous substrate

Vindiš, Tina (author), Blažič, Anja (author), Khayyat, Diaa (author), Potočnik, Tjaša (author), Sachdev, Shaurya (author), Rems, Lea (author)

Gene electrotransfer is one of the main non-viral methods for intracellular delivery of plasmid DNA, wherein pulsed electric fields are used to transiently permeabilize the cell membrane, allowing enhanced transmembrane transport. By localizing the electric field over small portions of the cell membrane using nanostructured substrates, it is possible to increase considerably the gene electrotransfer efficiency while preserving cell viability. In this study, we expand the frontier of localized electroporation by designing an electrotransfer approach based on commercially available cell culture inserts with polyethylene-terephthalate (PET) porous substrate. We first use multiscale numerical modeling to determine the pulse parameters, substrate pore size, and other factors that are expected to result in successful gene electrotransfer. Based on the numerical results, we design a simple device combining an insert with substrate containing pores with 0.4 µm or 1.0 µm diameter, a multiwell plate, and a pair of wire electrodes. We test the device in three mammalian cell lines and obtain transfection efficiencies similar to those achieved with conventional bulk electroporation, but at better cell viability and with low-voltage pulses that do not require the use of expensive electroporators. Our combined theoretical and experimental analysis calls for further systematic studies that will investigate the influence of substrate pore size and porosity on gene electrotransfer efficiency and cell viability.

• Language: English
• Keywords: localized electroporation, gene electrotransfer, plasmid, transfection, cell structure insert, numerical modeling, Chinese hamster ovary cells, myoblasts, C2C12 cell line, H9C2 cell line
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FE - Faculty of Electrical Engineering
• Publication status: Published
• Publication version: Version of Record
• Year: 2022
• Number of pages: 21 str.
• Numbering: Vol. 14, iss. 19, art. 1959
• PID: 20.500.12556/RUL-148018
• DOI: 10.3390/pharmaceutics14091959
• COBISS.SI-ID: 122088451
• Publication date in RUL: 20.07.2023

Record is a part of a journal

• Title: Pharmaceutics
• Shortened title: Pharmaceutics
• Publisher: MDPI
• ISSN: 1999-4923
• COBISS.SI-ID: 517949977

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: lokalizirana elektroporacija, genska elektrotransfekcija, transfekcija, vstavek za mikrotitrsko ploščo, numerično modeliranje, ovarijske celice kitajskega hrčka, celična linija C2C12, celična linija H9C2