title: A new spiral microelectrode assembly for electroporation and impedance measurements of adherent cell monolayers.
abstract: In this study, a new microelectrode assembly based on spiral geometry applicable to in situ electroporation of adherent cell monolayers on standard multiwell plates is presented. Furthermore, the structure is specially conceived to perform electrical impedance spectroscopy (EIS) measurements during electroporation. Its performance for cell membrane permeabilization is tested with a fluorescent probe. Gene electrotransfer is also assayed using a plasmid DNA encoding GFP in four different cell lines (CHO, HEK293, 3T3-L1 and FTO2B). Additionally, siRNA α-GFP electrotransfection is tested in GFP gene-expressing CHO cells. Our data show considerable differences between permeabilization and gene transfer results and cell line dependence on gene expression rates. Successful siRNA electro-mediated delivery is also achieved. We demonstrate the applicability of our device for electroporation-mediated gene transfer of adherent cells in standard laboratory conditions. Finally, electrical impedance measurements during electroporation of CHO and 3T3-L1 cells are also given.
authors.affiliation: Electronic and Biomedical Instrumentation Group, Department of Electrical Engineering, Universitat Politecnica de Catalunya, Barcelona, Spain, tomas.garcia.sanchez@upc.edu.
authors.forename: Tomás
authors.initials: T
authors.lastname: García-Sánchez
authors.forename: María
authors.initials: M
authors.lastname: Guitart
authors.forename: Javier
authors.initials: J
authors.lastname: Rosell-Ferrer
authors.forename: Anna M
authors.initials: AM
authors.lastname: Gómez-Foix
authors.forename: Ramon
authors.initials: R
authors.lastname: Bragós
mesh_terms: D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry; D002470:Cell Survival / Q000502:physiology; D003198:Computer Simulation; D003412:Cricetulus; D017097:Electric Impedance; D018274:Electroporation / Q000379:methods*; D004867:Equipment Design; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008839:Microelectrodes; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats
chemical_list: D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins