Electroporation – Advantages and Drawbacks for Delivery of Drug, Gene and Vaccine

Bolhassani, Azam; Khavari, Afshin; Orafa, Zahra

doi:10.5772/58376

Cited by 37 publications

(29 citation statements)

References 79 publications

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“…Engineering the genomes of primary human cells has significant therapeutic potential, but clinical translation is limited by efficacy and safety considerations associated with current delivery technologies ( 1 – 5 ). For example, advances in genome editing and gene therapy have brought hope for the development of new therapeutics in areas such as T cell engineering ( 6 ), hematopoietic stem cell (HSC) therapies ( 7 ), and regenerative medicine ( 8 ).…”

mentioning

confidence: 99%

Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo

DiTommaso

Cole

Cassereau

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

148

180

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SignificanceEx vivo manipulation of primary cells is critical to the success of this emerging generation of cell-based therapies, such as chimeric antigen receptor T cells for the treatment of cancer and CRISPR for the correction of developmental diseases. However, the limitations of existing delivery approaches may dramatically restrict the impact of genetic engineering to study and treat disease. In this paper, we compared electroporation to a microfluidic membrane deformation technique termed “squeezing” and found that squeezed cells had dramatically fewer side effects than electroporation and gene expression profiles similar to those of unmanipulated cells. The significant differences in outcomes from the two techniques underscores the importance of understanding the impact of intracellular delivery methods on cell function for research and clinical applications.

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mentioning

confidence: 99%

Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo

DiTommaso

Cole

Cassereau

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

148

180

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“…Electroporation is a non-viral technique used to deliver DNA, RNA, and proteins (including plasmid DNA (pDNA) vectors) to biological cells. Through the application of external electric fields of appropriate strength, duration, form, and number, a reversible increase in permeability is achieved to allow delivery of both small and large molecules through an otherwise impermeable cell membrane 8 . For many applications, electroporation is advantageous compared to viral-mediated gene delivery.…”

mentioning

confidence: 99%

“…Electroporation outcomes are typically defined as the resulting cell viability, defined as the percentage of living cells following electroporation compared to a non-electroporated control, and electro-transfection efficiency (eTE), defined as the percentage of cells receiving or expressing the delivered vector. These outcomes are dependent on a variety of experimental parameters including: electric pulse strength and duration, number of electric pulses applied, cell type, cell density, pDNA concentration, buffer conductivity, and buffer composition [8][9][10][11][12] . Not only does such a large number of experimental variables increase the complexity of protocol optimization, it has led to a vast landscape of published work, making it difficult to draw conclusions among them.…”

mentioning

confidence: 99%

The effects of electroporation buffer composition on cell viability and electro-transfection efficiency

Sherba

Hogquist

Lin

et al. 2020

Sci Rep

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Electroporation is an electro-physical, non-viral approach to perform DNA, RNA, and protein transfections of cells. Upon application of an electric field, the cell membrane is compromised, allowing the delivery of exogenous materials into cells. Cell viability and electro-transfection efficiency (eTE) are dependent on various experimental factors, including pulse waveform, vector concentration, cell type/density, and electroporation buffer properties. In this work, the effects of buffer composition on cell viability and eTE were systematically explored for plasmid DNA encoding green fluorescent protein following electroporation of 3T3 fibroblasts. A HEPES-based buffer was used in conjunction with various salts and sugars to modulate conductivity and osmolality, respectively. Pulse applications were chosen to maintain constant applied electrical energy (J) or total charge flux (C/m 2). The energy of the pulse application primarily dictated cell viability, with Mg 2+-based buffers expanding the reversible electroporation range. The enhancement of viability with Mg 2+-based buffers led to the hypothesis that this enhancement is due to ATPase activation via re-establishing ionic homeostasis. We show preliminary evidence for this mechanism by demonstrating that the enhanced viability is eliminated by introducing lidocaine, an ATPase inhibitor. However, Mg 2+ also hinders eTE compared to K +-based buffers. Collectively, the results demonstrate that the rational selection of pulsing conditions and buffer compositions are critical for the design of electroporation protocols to maximize viability and eTE.

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“…DEP of cancer cells has been widely researched and demonstrated in various applications including control of cell movements [1], electroporation [2], and killing of cells [3]. The advantages of using DEP is the label-free targeting of neutrally charged particles that can be sorted towards or away from regions of high electric field gradients in a non-uniform electric field region.…”

Section: Introductionmentioning

confidence: 99%

“…DEP of cancer cells has been widely researched and demonstrated in various applications including control of cell movements [1], electroporation [2], and killing of cells [3]. The shrinkage depending on their electrical properties, where these deformative occurrences may or may not result in the formation of reversible or irreversible micropores in the cell membrane.…”

Section: Introductionmentioning

confidence: 99%

Dielectrophoretic deformation of breast cancer cells for lab on a chip applications

et al. 2019

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This paper presents the development and experimental analysis of a curved microelectrode platform for the DEP deformation of breast cancer cells (MDA‐MB‐231). The platform is composed of arrays of curved DEP microelectrodes which are patterned onto a glass slide and samples containing MDA‐MB‐231 cells are pipetted onto the platform's surface. Finite element method is utilised to characterise the electric field gradient and DEP field. The performance of the system is assessed with MDA‐MB‐231 cells in a low conductivity 1% DMEM suspending medium. We applied sinusoidal wave AC potential at peak to peak voltages of 2, 5, and 10 Vpp at both 10 kHz and 50 MHz. We observed cell blebbing and cell shrinkage and analyzed the percentage of shrinkage of the cells. The experiments demonstrated higher percentage of cell shrinkage when cells are exposed to higher frequency and peak to peak voltage electric field.

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Electroporation – Advantages and Drawbacks for Delivery of Drug, Gene and Vaccine

Cited by 37 publications

References 79 publications

Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo

Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo

The effects of electroporation buffer composition on cell viability and electro-transfection efficiency

Dielectrophoretic deformation of breast cancer cells for lab on a chip applications

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