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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| Format: | Article |
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MDPI AG
2023-03-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/3/1004 |
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| author | Alexia de Caro Elisabeth Bellard Jelena Kolosnjaj-Tabi Muriel Golzio Marie-Pierre Rols |
| author_facet | Alexia de Caro Elisabeth Bellard Jelena Kolosnjaj-Tabi Muriel Golzio Marie-Pierre Rols |
| author_sort | Alexia de Caro |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-11T06:00:50Z |
| format | Article |
| id | doaj.art-09219703b2ed425c904bdcea9d2e643d |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T06:00:50Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-09219703b2ed425c904bdcea9d2e643d2023-11-17T13:17:27ZengMDPI AGPharmaceutics1999-49232023-03-01153100410.3390/pharmaceutics15031004Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative StudyAlexia de Caro0Elisabeth Bellard1Jelena Kolosnjaj-Tabi2Muriel Golzio3Marie-Pierre Rols4Institut de Pharmacologie et de Biologie Structurale du CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse CEDEX, FranceInstitut de Pharmacologie et de Biologie Structurale du CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse CEDEX, FranceInstitut de Pharmacologie et de Biologie Structurale du CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse CEDEX, FranceInstitut de Pharmacologie et de Biologie Structurale du CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse CEDEX, FranceInstitut de Pharmacologie et de Biologie Structurale du CNRS UMR 5089, 205, Route de Narbonne, 31077 Toulouse CEDEX, FranceElectroporation, 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.https://www.mdpi.com/1999-4923/15/3/1004electropermeabilizationgene electrotransfercells suspensionsspheroidsmicroscopyclearing |
| spellingShingle | Alexia de Caro Elisabeth Bellard Jelena Kolosnjaj-Tabi Muriel Golzio Marie-Pierre Rols Gene Electrotransfer Efficiency in 2D and 3D Cancer Cell Models Using Different Electroporation Protocols: A Comparative Study Pharmaceutics electropermeabilization gene electrotransfer cells suspensions spheroids microscopy clearing |
| 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 | electropermeabilization gene electrotransfer cells suspensions spheroids microscopy clearing |
| url | https://www.mdpi.com/1999-4923/15/3/1004 |
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