The dependence of efficiency of transmembrane molecular transfer using electroporation on medium viscosity

Sungailaitė, Sandra; Ruzgys, Paulius; Šatkauskienė, Ingrida; Čepurnienė, Karolina; Šatkauskas, Saulius

doi:10.1002/jgm.2825

Cited by 6 publications

(4 citation statements)

References 37 publications

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“…A recent study shows that, after using Ficoll to increase medium viscosity and so reduce DNA mobility and electrotransfer, the application of an LV pulse in addition to the HV one can recover to some extend the transfection efficiency. This reinforces the hypothesis of DNA electrophoresis being crucial for DNA electrotransfer [188]. This observation was confirmed both in vitro and in vivo for numerous tissues [29, 138, 189-195].…”

Section: Gene Electrotransfersupporting

confidence: 86%

Gene Electrotransfer: A Mechanistic Perspective

Rosazza¹,

Meglič²,

Zumbusch³

et al. 2016

CGT

181

142

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Gene electrotransfer is a powerful method of DNA delivery offering several medical applications, among the most promising of which are DNA vaccination and gene therapy for cancer treatment. Electroporation entails the application of electric fields to cells which then experience a local and transient change of membrane permeability. Although gene electrotransfer has been extensively studied in in vitro and in vivo environments, the mechanisms by which DNA enters and navigates through cells are not fully understood. Here we present a comprehensive review of the body of knowledge concerning gene electrotransfer that has been accumulated over the last three decades. For that purpose, after briefly reviewing the medical applications that gene electrotransfer can provide, we outline membrane electropermeabilization, a key process for the delivery of DNA and smaller molecules. Since gene electrotransfer is a multipart process, we proceed our review in describing step by step our current understanding, with particular emphasis on DNA internalization and intracellular trafficking. Finally, we turn our attention to in vivo testing and methodology for gene electrotransfer.

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Section: Gene Electrotransfersupporting

confidence: 86%

Gene Electrotransfer: A Mechanistic Perspective

Rosazza¹,

Meglič²,

Zumbusch³

et al. 2016

CGT

181

142

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“…The previous research on electroporation optimization focused mostly on the empirical determination of input parameters that describe electric field or single characteristics of the medium [31]. However, the relationship between the input parameters and the therapeutic outcome is indirect, as the final state of the system is achieved over time due to the redistribution of the constituents induced by biophysical processes.…”

Section: Discussionmentioning

confidence: 99%

Investigation of Plasmid DNA Delivery and Cell Viability Dynamics for Optimal Cell Electrotransfection In Vitro

Chopra¹,

Ruzgys²,

Maciulevičius³

et al. 2020

Applied Sciences

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Electroporation is an effective method for delivering plasmid DNA molecules into cells. The efficiency of gene electrotransfer depends on several factors. To achieve high transfection efficiency while maintaining cell viability is a tedious task in electroporation. Here, we present a combined study in which the dynamics of both evaluation types of transfection efficiency and the cell viability were evaluated in dependence of plasmid concentration as well as at the different number of high voltage (HV) electric pulses. The results of this study reveal a quantitative sigmoidal (R2 > 0.95) dependence of the transfection efficiency and cell viability on the distance between the cell membrane and the nearest plasmid. We propose this distance value as a new, more accurate output parameter that could be used in further optimization studies as a predictor and a measure of electrotransfection efficiency.

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“…In the specific case of PI, it has been suggested that extracellular conductivity might have an influence not only of permeabilisation but also on the electrophoretic transport of propidium since it can be enhanced at low conductivity 32 . In the case of bleomycin, no influence of the conductivity on the transport is expected since transport occurs almost exclusively by diffusion and is not transported by electrophoresis 33 . However, a thorough study using different markers would re-inforce the present data and enable to draw larger conclusion regarding the impact of extracellular conductivity on permeabilisation.…”

Section: Discussionmentioning

confidence: 99%

Impact of external medium conductivity on cell membrane electropermeabilization by microsecond and nanosecond electric pulses

Silve

Leray

Poignard

et al. 2016

Sci Rep

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The impact of external medium conductivity on the efficiency of the reversible permeabilisation caused by pulsed electric fields was investigated. Pulses of 12 ns, 102 ns or 100 μs were investigated. Whenever permeabilisation could be detected after the delivery of one single pulse, media of lower conductivity induced more efficient reversible permeabilisation and thus independently of the medium composition. Effect of medium conductivity can however be hidden by some saturation effects, for example when pulses are cumulated (use of trains of 8 pulses) or when the detection method is not sensitive enough. This explains the contradicting results that can be found in the literature. The new data are complementary to those of one of our previous study in which an opposite effect of the conductivity was highlighted. It stresses that the conductivity of the medium influences the reversible permeabilization by several ways. Moreover, these results clearly indicate that electropermeabilisation does not linearly depend on the energy delivered to the cells.

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The dependence of efficiency of transmembrane molecular transfer using electroporation on medium viscosity

Abstract: These findings demonstrate that transfection is very susceptible to medium viscosity and highlights the importance of the electrophoretic component in experiments when a considerable transfection level is needed.

Cited by 6 publications

References 37 publications

Gene Electrotransfer: A Mechanistic Perspective

Gene Electrotransfer: A Mechanistic Perspective

Investigation of Plasmid DNA Delivery and Cell Viability Dynamics for Optimal Cell Electrotransfection In Vitro

Impact of external medium conductivity on cell membrane electropermeabilization by microsecond and nanosecond electric pulses

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