Mechanisms involved in gene electrotransfer using high- and low-voltage pulses — An in vitro study

Kandušer, Maša; Miklavčič, Damijan; Pavlin, Mojca

doi:10.1016/j.bioelechem.2008.09.002

Cited by 117 publications

(118 citation statements)

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“…After an HV pulse, all cells are permeabilized and the LV pulse brings plasmids into contact with cells due to electrophoretic forces. This double-pulse method was shown to be valid in vitro when using suboptimal plasmid concentration (Kandušer et al 2009). In the present study we observed the effect of electric field orientation using HVLV pulse trains with different modulations of the pulse polarities.…”

Section: Introductionmentioning

confidence: 94%

Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses

et al. 2012

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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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Section: Introductionmentioning

confidence: 94%

Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses

et al. 2012

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“…For example, for the introduction of small molecules, pulses with amplitudes in the range of 1 kV/cm and durations extending from hundred μs to ms are used [11]- [17]. Larger molecules can be introduced using three different combinations of pulse parameters: 1) with pulse amplitudes up to few kV/cm, lasting from few μs to hundred μs [18], [19]; 2) with low pulse amplitudes of few hundred V/cm but durations ranging into tens of ms [20]; 3) with a combination of short high-amplitude pulses and long lowamplitude pulses (mostly for the uptake of DNA) [21]- [25]. For sterilization in food and drink industry, the pulse amplitudes should be larger than 15 kV/cm in order to electroporate the membranes of microorganisms, which are smaller than eukaryotic cells, while pulse durations range from μs to ms [9], [26].…”

Section: Introductionmentioning

confidence: 99%

Equivalent Pulse Parameters for Electroporation

Pucihar

Krmelj

Reberšek

et al. 2011

IEEE Trans. Biomed. Eng.

192

109

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Abstract-Electroporation-based applications require the use of specific pulse parameters for a successful outcome. When recommended values of pulse parameters cannot be set, similar outcomes can be obtained by using equivalent pulse parameters. We determined the relations between the amplitude and duration/number of pulses resulting in the same fraction of electroporated cells. Pulse duration was varied from 150 ns to 100 ms, and the number of pulses from 1 to 128. Fura 2-AM was used to determine electroporation of cells to Ca 2+ . With longer pulses or higher number of pulses, lower amplitudes are needed for the same fraction of electroporated cells. The expression derived from the model of electroporation could describe the measured data on the whole interval of pulse durations. In a narrower range (0.1-100 ms), less complex, logarithmic or power functions could be used instead. The relation between amplitude and number of pulses could best be described with a power function or an exponential function. We show that relatively simple two-parameter power or logarithmic functions are useful when equivalent pulse parameters for electroporation are sought. Such mathematical relations between pulse parameters can be important in planning of electroporationbased treatments, such as electrochemotherapy and nonthermal irreversible electroporation.

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“…In EGT the electric pulses are used to electropermeabilize the cell membranes (as in ECT) and also to facilitate the electrophoretic transfer of extracellular genetic material into the cells [28][29][30][31][32][33][34][35]. The potential of EGT for local gene therapy without the use of viral vectors has been demonstrated in vivo and in a clinical study for tissues, including muscle, skin, liver and tumors [31,34,[36][37][38][39][40].…”

mentioning

confidence: 99%

Antivascular effects of electrochemotherapy: implications in treatment of bleeding metastases

Jarm

Čemažar

Miklavčič

et al. 2010

Expert Review of Anticancer Therapy

200

132

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Mechanisms involved in gene electrotransfer using high- and low-voltage pulses — An in vitro study

Cited by 117 publications

References 50 publications

Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses

Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses

Equivalent Pulse Parameters for Electroporation

Antivascular effects of electrochemotherapy: implications in treatment of bleeding metastases

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