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A new spiral microelectrode assembly for electroporation and impedance measurements of adherent cell monolayers

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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.

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Acknowledgements

This study is supported by grants SAF2009-07559 and SAF2012-37480 from the Spanish Ministerio de Ciencia e Innovación (MCI) and CIBERDEM de Diabetes y Enfermedades Metabólicas Asociadas (CB07/08/0012). We also thank Anna Orozco and Alfonso Mendez for their technical support.

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Authors and Affiliations

  1. Electronic and Biomedical Instrumentation Group, Department of Electrical Engineering, Universitat Politecnica de Catalunya, Barcelona, Spain

    Tomás García-Sánchez, Javier Rosell-Ferrer & Ramon Bragós

  2. Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, IBUB, Universitat de Barcelona (UB), Barcelona, Spain

    María Guitart & Anna M. Gómez-Foix

  3. CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Instituto de Salud Carlos III, Madrid, Spain

    Anna M. Gómez-Foix

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  1. Tomás García-Sánchez
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  2. María Guitart
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  3. Javier Rosell-Ferrer
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  4. Anna M. Gómez-Foix
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  5. Ramon Bragós
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Correspondence to Tomás García-Sánchez.

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García-Sánchez, T., Guitart, M., Rosell-Ferrer, J. et al. A new spiral microelectrode assembly for electroporation and impedance measurements of adherent cell monolayers. Biomed Microdevices 16, 575–590 (2014). https://doi.org/10.1007/s10544-014-9860-6

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