Abstract
Electropermeabilization is a biological physical process in response to the presence of an applied electric field that is used for the transfer of hydrophilic molecules such as anticancer drugs or DNA across the plasma membranes of living cells. The molecular processes that support the transfer are poorly known. The aim of our study was to investigate the effect of high-voltage and low-voltage (HVLV) pulses in vitro with different orientations on cell permeabilization, viability and gene transfection. We monitored the permeabilization with unipolar and bipolar HVLV pulses with different train repetition pulses, showing that HVLV pulses increase cell permeabilization and cell viability. Gene transfer was also observed by measuring green fluorescent protein (GFP) expression. The expression was the same for HVLV pulses and electrogenotherapy pulses for in vitro experimentation. As the viability was better preserved for HVLV-pulsed cells, we managed to increase the number of GFP-expressing cells by up to 65 % under this condition. The use of bipolar HVLV train pulses increased gene expression to a higher extent, probably by affecting a larger part of the cell surface.
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Acknowledgments
This work was supported by the European Union project OncomiR (Grant 201102). The authors thank Betatech for the joint development of generators and the TRI (Toulouse Réseau d’Imagerie) platform at the IPBS (funded by the region Midi-Pyrénées, the Communauté du Grand Toulouse and the FEDER) for providing access to the flow cytometer.
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Orio, J., Coustets, M., Mauroy, C. et al. Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses. J Membrane Biol 245, 661–666 (2012). https://doi.org/10.1007/s00232-012-9475-x
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DOI: https://doi.org/10.1007/s00232-012-9475-x