A 3D in vitro spheroid model as a way to study the mechanisms of electroporation
Abstract
Electropermeabilization is a physical method to deliver molecules into cells and tissues. Clinical applications have been successfully developed for antitumoral drug delivery and clinical trials for gene electrotransfer are currently underway. However, little is known about the mechanisms involved in this transfer. The main difficulties stem from the lack of single cell models which reliably replicate the complex in vivo environment. In order to increase our understanding of the DNA electrotransfer process, we exploited multicellular tumor spheroids as an ex vivo model of tumor. We used confocal microscopy to visualize the repartition of permeabilized cells in spheroids subjected to electric pulses. Our results reveal that even if cells can be efficiently permeabilized with electric fields, including those cells present inside the spheroids, gene expression is by contrast limited to the external layers of cells. Taken together, these results, in agreement with the ones obtained in tumors, indicate that the spheroid model is more relevant to an in vivo situation than cells cultured as monolayers. They validate the spheroid model as a way to study electro-mediated gene delivery processes.
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Transfected cells were heterogeneously distributed and appeared to be only localized in the outer layers of the spheroids.
“A striking observation is that transfected cells were heterogeneously distributed and appeared to be only localized in the outer layers of the spheroids.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [28]
These results suggest that limited access to central regions of MCTS remains a significant barrier to gene delivery.
“These results suggest that limited access to central regions of MCTS remains a significant barrier to gene delivery.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [28]
Uptake efficiency of macromolecules such as plasmid DNA was inefficient; only a few cells from colon cancer MCTS were transfected using a plasmid coding for GFP.
“However, the uptake efficiency of macromolecules such as plasmid DNA was found to be inefficient. Indeed, only a few cells from colon cancer MCTS were transfected using a plasmid that codes for the GFP.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [28]
No difference was detected between cells at the periphery and cells located deeper within the spheroid's core in that electroporation study.
“In addition, no difference was detected between cells at the periphery and the cells located deeper within the spheroid's core.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [28]
MCTS from mouse sarcoma were transfected via electroporation in the presence of propidium iodide; practically the entire cell population was permeabilized, as measured by propidium iodide uptake.
“In another study, MCTS from mouse sarcoma were transfected via electroporation in the presence of propidium iodide. The method was highly efficient since practically the entire cell population was permeabilized, as measured by propidium iodide uptake (28).”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [28]
Cites
Canatella, P. J., Black, M. M., Bonnichsen, D. M., McKenna, C., & Prausnitz, M. R. (2004) · Biophysical Journal · doi:10.1016/s0006-3495(04)74374-x · Cited by 89 · see citing works
Optimising non-viral gene delivery in a tumour spheroid model
Mellor, H. R., Davies, L. A., Caspar, H., Pringle, C. R., Hyde, S. C., Gill, D. R., & Callaghan, R. (2006) · The Journal of Gene Medicine · doi:10.1002/jgm.947 · Cited by 49 · see citing works
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Laperrousaz, B., Porte, S., Gerbaud, S., Härmä, V., Kermarrec, F., Hourtane, V., Bottausci, F., Gidrol, X., & Picollet-D’hahan, N. (2018) · Nucleic Acids Research · doi:10.1093/nar/gky030 · Cited by 54 · see citing works
“Even in spheroid models produced by cell aggregation and devoid of ECM, direct 3D transfection is usually limited to the outer layer of cells (28), which reduces its relevance and leads to heterogeneous results.”
BackgroundMethod
- Authors
- Wasungu, L., Escoffre, J.-M., Valette, A., Teissie, J., & Rols, M.-P.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Corresponding author
- Rols, M.-P.
- Venue
- International Journal of Pharmaceutics
- Publisher
- Elsevier BV
- Published
- 2009-09
- Volume
- 379
- Issue
- 2
- Pages
- 278-284
- ISSN
- 0378-5173 (ISSN-L)
- OpenAlex
- W2061427203
- Cited by
- 58
- References
- 2
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 1 · 2023: 4 · 2022: 2 · 2021: 6 · 2020: 3 · 2019: 3 · 2018: 2 · 2017: 4 · 2016: 5 · 2015: 6 · 2014: 2 · 2013: 4 · 2012: 2
- FWCI
- 3.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- International Journal of Pharmaceutics (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Microbial Inactivation Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Engineering › Biomedical Engineering › Ultrasound and Hyperthermia Applications
- Keywords
- electroporation, tumor spheroids, gene electrotransfer, DNA delivery, electric field effects on cells, in vitro tumor model, electric pulse treatment, anticancer drug delivery
- MeSH terms
- Animals, Humans, Mice, Inbred C57BL, Models, Molecular, Molecular Conformation, Permeability, Gene Transfer Techniques, Electroporation, Spheroids, Cellular, HCT116 Cells, Mice
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