Elsevier

Available online 25 January 2020, 111877

Experimental Cell Research

Highlights

Reversible electroporation was used for delivery of large conjugates into the cell.

msPEFs can be utilised to deliver stable antibodies into the cytosol.

msPEFs method allowed detection of IL-6 in viable cells.

Abstract

Reversible electroporation is the temporary permeabilization of cell membrane through the formation of transient pores created by short high voltage electric pulses. This method has numerous applications in biology and biotechnology and has become an important technique in molecular medicine. Reversible electroporation is usually used to transfer macromolecules into the cells. However, the delivery of large molecules such as proteins into cells without loss of cell viability remains a challenge. In our study, we investigated whether electroporation can be used for this purpose. The study was performed with the primary mouse splenocytes and Jurkat cell line. The electroporation efficacy was evaluated by flow cytometry. We used the reversible electroporation for intracellular marker detection investigating antibody and fluorescein-conjugated dextran transfer efficiency, cell viability and metabolic activity. We have found that reversible electroporation parameters can be optimized for efficient transfer of large molecules such as antibodies/proteins into live cells without a significant loss of cell viability. We conclude that a well-established and relatively easy method of reversible electroporation can be adjusted to detect intracellular biomarkers in viable cells. This is a new approach on how electroporation could be utilised in medicine and biological research to detect rare subpopulations of cells that produce specific markers and to keep cells viable. This would allow the use of these rare subpopulations of isolated cells for further research and personalized medicine.

Keywords

Electroporation

Antibody

Viability

Biomarker

IL-6

Splenocyte

Actin

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© 2020 Published by Elsevier Inc.



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