Tina Cimperman (Avtor), Anja Blažič (Avtor), Diaa Khayyat (Avtor), Tjaša Potočnik (Avtor), Shaurya Sachdev (Avtor), Lea Rems (Avtor)

Povzetek

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.

Ključne besede

lokalizirana elektroporacija;genska elektrotransfekcija;transfekcija;vstavek za mikrotitrsko ploščo;numerično modeliranje;ovarijske celice kitajskega hrčka;celična linija C2C12;celična linija H9C2;localized electroporation;gene electrotransfer;plasmid;transfection;cell structure insert;numerical modeling;Chinese hamster ovary cells;myoblasts;C2C12 cell line;H9C2 cell line;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: UL FE - Fakulteta za elektrotehniko
UDK: 602.621:577
COBISS: 122088451 Povezava se bo odprla v novem oknu
ISSN: 1999-4923
Št. ogledov: 70
Št. prenosov: 24
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarne ključne besede: lokalizirana elektroporacija;genska elektrotransfekcija;transfekcija;vstavek za mikrotitrsko ploščo;numerično modeliranje;ovarijske celice kitajskega hrčka;celična linija C2C12;celična linija H9C2;
Vrsta dela (COBISS): Članek v reviji
Strani: str. 1-21
Letnik: ǂiss. ǂ9
Zvezek: 1959
Čas izdaje: Sep. 2022
DOI: 10.3390/pharmaceutics14091959
ID: 19638995