In Vitro and Ex Vivo Gene Delivery to Cells by Electroporation

Hui, Sek Wen; Li, Lin Hong

doi:10.1385/1-59259-080-2:157

Cited by 14 publications

(19 citation statements)

References 31 publications

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“…Increased permeability of the cell membrane (indicated by dye uptake) occurs progressively longer after pulse application as the pulse duration shortens, and this temporal separation from the pulse application suggests that the membrane permeability effects are more likely due to alterations of cellular function rather than direct effects on surface membrane structure. The increase in cell size seen with pulses longer than 60 ns likely represents colloid osmotic swelling of cells in physiologic buffer, and as such, would be typical of electroporation with delayed resealing of the pores (Hui and Li, 2000).…”

Section: Discussionmentioning

confidence: 99%

The Effects of Intense Submicrosecond Electrical Pulses on Cells

Deng

Schoenbach

Buescher

et al. 2003

Biophysical Journal

218

129

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A simple electrical model for living cells predicts an increasing probability for electric field interactions with intracellular substructures of both prokaryotic and eukaryotic cells when the electric pulse duration is reduced into the sub-microsecond range. The validity of this hypothesis was verified experimentally by applying electrical pulses (durations 100 micros-60 ns, electric field intensities 3-150 kV/cm) to Jurkat cells suspended in physiologic buffer containing propidium iodide. Effects on Jurkat cells were assessed by means of temporally resolved fluorescence and light microscopy. For the longest applied pulses, immediate uptake of propidium iodide occurred consistent with electroporation as the cause of increased surface membrane permeability. For nanosecond pulses, more delayed propidium iodide uptake occurred with significantly later uptake of propidium iodide occurring after 60 ns pulses compared to 300 ns pulses. Cellular swelling occurred rapidly following 300 ns pulses, but was minimal following 60 ns pulses. These data indicate that submicrosecond pulses achieve temporally distinct effects on living cells compared to microsecond pulses. The longer pulses result in rapid permeability changes in the surface membrane that are relatively homogeneous across the cell population, consistent with electroporation, while shorter pulses cause surface membrane permeability changes that are temporally delayed and heterogeneous in their magnitude.

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Section: Discussionmentioning

confidence: 99%

The Effects of Intense Submicrosecond Electrical Pulses on Cells

Deng

Schoenbach

Buescher

et al. 2003

Biophysical Journal

218

129

View full text Add to dashboard Cite

show abstract

“…Interestingly, cells can survive up to 50% volume increase and still recover 429,430,435,437,445,449 . Above that, the risk of instant death from bursting becomes imminent 450 . It is known that swelling activates specific stress signaling events 451 and is a classic hallmark associated with necrotic cell death 452,453 .…”

Section: Membrane Disruption-mediated Delivery: Background Conceptsmentioning

confidence: 99%

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

2018

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Intracellular delivery is a key step in biological research and has enabled decades of biomedical discoveries. It is also becoming increasingly important in industrial and medical applications ranging from biomanufacture to cell-based therapies. Here, we review techniques for membrane disruption-based intracellular delivery from 1911 until the present. These methods achieve rapid, direct, and universal delivery of almost any cargo molecule or material that can be dispersed in solution. We start by covering the motivations for intracellular delivery and the challenges associated with the different cargo types-small molecules, proteins/peptides, nucleic acids, synthetic nanomaterials, and large cargo. The review then presents a broad comparison of delivery strategies followed by an analysis of membrane disruption mechanisms and the biology of the cell response. We cover mechanical, electrical, thermal, optical, and chemical strategies of membrane disruption with a particular emphasis on their applications and challenges to implementation. Throughout, we highlight specific mechanisms of membrane disruption and suggest areas in need of further experimentation. We hope the concepts discussed in our review inspire scientists and engineers with further ideas to improve intracellular delivery.

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“…In addition, it is expected that electric pulses will act to porate cells in vivo, leading to increased cellular uptake of the photosensitizer. A number of studies have demonstrated that electroporation (cell electropermeabilization, electrochemotherapy) is an important development for the in vitro (51) or in vivo (52,53) cellular delivery of a number of molecules. Because ALA–PDT relies on the intracellular conversion of ALA to PpIX, this is particularly critical.…”

Section: Discussionmentioning

confidence: 99%

Electrically Enhanced Percutaneous Delivery Of δ-Aminolevulinic Acid Using Electric Pulses and a DC Potential¶

Johnson

Hui

Oseroff

2007

Photochemistry and Photobiology

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Selectivity of photodynamic therapy can be improved with localized photosensitizer delivery, but topical administration is restricted by poor diffusion across the stratum corneum. We used electric pulses to increase transdermal transport of δ‐aminolevulinic acid (ALA), a precursor to the photosensitizer protoporphyrin IX (PpIX). ALA‐filled electrodes were attached to the surface of excised porcine skin or the dorsal surface of mice. Pulses were administered and, in some in vivo cases, a continuous DC potential (6 V) was concomitantly applied. For in vitro14C ALA penetration, 10 μm layers parallel to the stratum corneum were assayed by liquid scintillation analysis, and 10 μm cross sections were examined autoradiographically. As the electrical dose (voltage × frequency × pulse width × treatment duration) increased, there was an increase in penetration depth. In vivo delivery was assayed by measuring the fluorescence of PpIX in skin samples. A greater than two‐fold enhancement of PpIX production with electroporative delivery was seen versus that obtained with passive delivery. Superimposition of a DC potential resulted in a nearly three‐fold enhancement of PpIX production versus passive delivery. Levels were higher than the sum of PpIX detected after pulse‐alone and DC‐alone delivery. Electroporation and electrophoresis are likely factors in electrically enhanced delivery.

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In Vitro and Ex Vivo Gene Delivery to Cells by Electroporation

Cited by 14 publications

References 31 publications

The Effects of Intense Submicrosecond Electrical Pulses on Cells

The Effects of Intense Submicrosecond Electrical Pulses on Cells

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

Electrically Enhanced Percutaneous Delivery Of δ-Aminolevulinic Acid Using Electric Pulses and a DC Potential¶

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