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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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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. |
format | Online Article Text |
id | pubmed-10053976 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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