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Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study

Electroporation, a method relying on a pulsed electric field to induce transient cell membrane permeabilization, can be used as a non-viral method to transfer genes in vitro and in vivo. Such transfer holds great promise for cancer treatment, as it can induce or replace missing or non-functioning ge...

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Autores principales: de Caro, Alexia, Bellard, Elisabeth, Kolosnjaj-Tabi, Jelena, Golzio, Muriel, Rols, Marie-Pierre
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10053976/
https://www.ncbi.nlm.nih.gov/pubmed/36986866
http://dx.doi.org/10.3390/pharmaceutics15031004
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author de Caro, Alexia
Bellard, Elisabeth
Kolosnjaj-Tabi, Jelena
Golzio, Muriel
Rols, Marie-Pierre
author_facet de Caro, Alexia
Bellard, Elisabeth
Kolosnjaj-Tabi, Jelena
Golzio, Muriel
Rols, Marie-Pierre
author_sort de Caro, Alexia
collection PubMed
description Electroporation, a method relying on a pulsed electric field to induce transient cell membrane permeabilization, can be used as a non-viral method to transfer genes in vitro and in vivo. Such transfer holds great promise for cancer treatment, as it can induce or replace missing or non-functioning genes. Yet, while efficient in vitro, gene-electrotherapy remains challenging in tumors. To assess the differences of gene electrotransfer in respect to applied pulses in multi-dimensional (2D, 3D) cellular organizations, we herein compared pulsed electric field protocols applicable to electrochemotherapy and gene electrotherapy and different “High Voltage–Low Voltage” pulses. Our results show that all protocols can result in efficient permeabilization of 2D- and 3D-grown cells. However, their efficiency for gene delivery varies. The gene-electrotherapy protocol is the most efficient in cell suspensions, with a transfection rate of about 50%. Conversely, despite homogenous permeabilization of the entire 3D structure, none of the tested protocols allowed gene delivery beyond the rims of multicellular spheroids. Taken together, our findings highlight the importance of electric field intensity and the occurrence of cell permeabilization, and underline the significance of pulses’ duration, impacting plasmids’ electrophoretic drag. The latter is sterically hindered in 3D structures and prevents the delivery of genes into spheroids’ core.
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spelling pubmed-100539762023-03-30 Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study de Caro, Alexia Bellard, Elisabeth Kolosnjaj-Tabi, Jelena Golzio, Muriel Rols, Marie-Pierre Pharmaceutics Article Electroporation, a method relying on a pulsed electric field to induce transient cell membrane permeabilization, can be used as a non-viral method to transfer genes in vitro and in vivo. Such transfer holds great promise for cancer treatment, as it can induce or replace missing or non-functioning genes. Yet, while efficient in vitro, gene-electrotherapy remains challenging in tumors. To assess the differences of gene electrotransfer in respect to applied pulses in multi-dimensional (2D, 3D) cellular organizations, we herein compared pulsed electric field protocols applicable to electrochemotherapy and gene electrotherapy and different “High Voltage–Low Voltage” pulses. Our results show that all protocols can result in efficient permeabilization of 2D- and 3D-grown cells. However, their efficiency for gene delivery varies. The gene-electrotherapy protocol is the most efficient in cell suspensions, with a transfection rate of about 50%. Conversely, despite homogenous permeabilization of the entire 3D structure, none of the tested protocols allowed gene delivery beyond the rims of multicellular spheroids. Taken together, our findings highlight the importance of electric field intensity and the occurrence of cell permeabilization, and underline the significance of pulses’ duration, impacting plasmids’ electrophoretic drag. The latter is sterically hindered in 3D structures and prevents the delivery of genes into spheroids’ core. MDPI 2023-03-21 /pmc/articles/PMC10053976/ /pubmed/36986866 http://dx.doi.org/10.3390/pharmaceutics15031004 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
de Caro, Alexia
Bellard, Elisabeth
Kolosnjaj-Tabi, Jelena
Golzio, Muriel
Rols, Marie-Pierre
Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title_full Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title_fullStr Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title_full_unstemmed Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title_short Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study
title_sort gene electrotransfer efficiency in 2d and 3d cancer cell models using different electroporation protocols: a comparative study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10053976/
https://www.ncbi.nlm.nih.gov/pubmed/36986866
http://dx.doi.org/10.3390/pharmaceutics15031004
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