1. Interplay between Electric Field Strength and Number of Short-Duration Pulses for Efficient Gene Electrotransfer.
- Author
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Urbanskas, Ernestas, Jakštys, Baltramiejus, Venckus, Justinas, Malakauskaitė, Paulina, Šatkauskienė, Ingrida, Morkvėnaitė-Vilkončienė, Inga, and Šatkauskas, Saulius
- Subjects
CHO cell ,SQUARE waves ,GENE expression ,CELL survival ,ELECTRIC fields ,ELECTROPORATION - Abstract
Electroporation is a method that shows great promise as a non-viral approach for delivering genes by using high-voltage electric pulses to introduce DNA into cells to induce transient gene expression. This research aimed to evaluate the interplay between electric pulse intensity and 100 µs-duration pulse numbers as an outcome of gene electrotransfer efficacy and cell viability. Our results indicated a close relationship between pulse number and electric field strength regarding gene electrotransfer efficacy; higher electric pulse intensity resulted in fewer pulses needed to achieve the same gene electrotransfer efficacy. Subsequently, an increase in pulse number had a more negative impact on overall gene electrotransfer by significantly reducing cell viability. Based on our data, the best pulse parameters to transfect CHO cells with the pMax-GFP plasmid were using 5 HV square wave pulses of 1000 V/cm and 2 HV of 1600 V/cm, correspondingly resulting in 55 and 71% of transfected cells and maintaining 79 and 54% proliferating cells. This shows ESOPE-like 100 µs-duration pulse protocols can be used simultaneously to deliver cytotoxic drugs as well as immune response regulating genetically encoded cytokines. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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