Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells
Abstract Electric pulses used in electroporation-based treatments have been shown to affect the excitability of muscle and neuronal cells. However, understanding the interplay between electroporation and electrophysiological response of excitable cells is complex, since both ion channel gating and e...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-01-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-51073-5 |
| _version_ | 1827388540855517184 |
|---|---|
| author | Tina Batista Napotnik Bor Kos Tomaž Jarm Damijan Miklavčič Rodney P. O’Connor Lea Rems |
| author_facet | Tina Batista Napotnik Bor Kos Tomaž Jarm Damijan Miklavčič Rodney P. O’Connor Lea Rems |
| author_sort | Tina Batista Napotnik |
| collection | DOAJ |
| description | Abstract Electric pulses used in electroporation-based treatments have been shown to affect the excitability of muscle and neuronal cells. However, understanding the interplay between electroporation and electrophysiological response of excitable cells is complex, since both ion channel gating and electroporation depend on dynamic changes in the transmembrane voltage (TMV). In this study, a genetically engineered human embryonic kidney cells expressing NaV1.5 and Kir2.1, a minimal complementary channels required for excitability (named S-HEK), was characterized as a simple cell model used for studying the effects of electroporation in excitable cells. S-HEK cells and their non-excitable counterparts (NS-HEK) were exposed to 100 µs pulses of increasing electric field strength. Changes in TMV, plasma membrane permeability, and intracellular Ca2+ were monitored with fluorescence microscopy. We found that a very mild electroporation, undetectable with the classical propidium assay but associated with a transient increase in intracellular Ca2+, can already have a profound effect on excitability close to the electrostimulation threshold, as corroborated by multiscale computational modelling. These results are of great relevance for understanding the effects of pulse delivery on cell excitability observed in context of the rapidly developing cardiac pulsed field ablation as well as other electroporation-based treatments in excitable tissues. |
| first_indexed | 2024-03-08T16:20:19Z |
| format | Article |
| id | doaj.art-4736a2f599ba4092b5049d7f64e4f49d |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-08T16:20:19Z |
| publishDate | 2024-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-4736a2f599ba4092b5049d7f64e4f49d2024-01-07T12:23:44ZengNature PortfolioScientific Reports2045-23222024-01-0114111810.1038/s41598-023-51073-5Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cellsTina Batista Napotnik0Bor Kos1Tomaž Jarm2Damijan Miklavčič3Rodney P. O’Connor4Lea Rems5University of Ljubljana, Faculty of Electrical EngineeringUniversity of Ljubljana, Faculty of Electrical EngineeringUniversity of Ljubljana, Faculty of Electrical EngineeringUniversity of Ljubljana, Faculty of Electrical EngineeringÉcole des Mines de Saint-Étienne, Department of Bioelectronics, Georges Charpak Campus, Centre Microélectronique de ProvenceUniversity of Ljubljana, Faculty of Electrical EngineeringAbstract Electric pulses used in electroporation-based treatments have been shown to affect the excitability of muscle and neuronal cells. However, understanding the interplay between electroporation and electrophysiological response of excitable cells is complex, since both ion channel gating and electroporation depend on dynamic changes in the transmembrane voltage (TMV). In this study, a genetically engineered human embryonic kidney cells expressing NaV1.5 and Kir2.1, a minimal complementary channels required for excitability (named S-HEK), was characterized as a simple cell model used for studying the effects of electroporation in excitable cells. S-HEK cells and their non-excitable counterparts (NS-HEK) were exposed to 100 µs pulses of increasing electric field strength. Changes in TMV, plasma membrane permeability, and intracellular Ca2+ were monitored with fluorescence microscopy. We found that a very mild electroporation, undetectable with the classical propidium assay but associated with a transient increase in intracellular Ca2+, can already have a profound effect on excitability close to the electrostimulation threshold, as corroborated by multiscale computational modelling. These results are of great relevance for understanding the effects of pulse delivery on cell excitability observed in context of the rapidly developing cardiac pulsed field ablation as well as other electroporation-based treatments in excitable tissues.https://doi.org/10.1038/s41598-023-51073-5 |
| spellingShingle | Tina Batista Napotnik Bor Kos Tomaž Jarm Damijan Miklavčič Rodney P. O’Connor Lea Rems Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells Scientific Reports |
| title | Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells |
| title_full | Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells |
| title_fullStr | Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells |
| title_full_unstemmed | Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells |
| title_short | Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells |
| title_sort | genetically engineered hek cells as a valuable tool for studying electroporation in excitable cells |
| url | https://doi.org/10.1038/s41598-023-51073-5 |
| work_keys_str_mv | AT tinabatistanapotnik geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells AT borkos geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells AT tomazjarm geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells AT damijanmiklavcic geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells AT rodneypoconnor geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells AT learems geneticallyengineeredhekcellsasavaluabletoolforstudyingelectroporationinexcitablecells |