Abstract
High-voltage pulses applied to a cell suspension cause not only cell membrane permeabilization, but a variety of electrolysis reactions to also occur at the electrode–solution interfaces. Here, the cytotoxicity of a culture medium treated by a single electric pulse and the role of the iron ions in this cytotoxicity were studied in vitro. The experiments were carried out on mouse hepatoma MH-22A, rat glioma C6, and Chinese hamster ovary cells. The cell culture medium treated with a high-voltage pulse was highly cytotoxic. All cells died in the medium treated by a single electric pulse with a duration of 2 ms and an amplitude of just 0.2 kV/cm. The medium treated with a shorter pulse was less cytotoxic. The cell viability was inversely proportional to the amount of electric charge that flowed through the solution. The amount of iron ions released from the stainless steel anode (>0.5 mM) was enough to reduce cell viability. However, iron ions were not the sole reason of cell death. To kill all MH-22A and CHO cells, the concentration of Fe3+ ions in a medium of more than 2 mM was required.
Highlights
Electroporation is an effective method for the modification of cell membrane permeability, the number of applications of which are steadily increasing in biology, oncology, genetics, immunology, and biotechnology [1,2]
Several studies reported that media treated with high-voltage electric pulses, which are usually utilized for cell electroporation or other electromanipulation purposes, demonstrate cytotoxic and bactericidal features [38,39,40]
On the basis of the results obtained in the present study on the cytotoxicity of a cell culture medium pre-treated with a high-voltage pulse and iron (Fe2+ & Fe3+) ions, the following main conclusions can be drawn: Products of electrolysis reactions can affect cell viability—a cell culture medium treated by an electric pulse is highly cytotoxic depending on the electrode material
Summary
Electroporation is an effective method for the modification of cell membrane permeability, the number of applications of which are steadily increasing in biology, oncology, genetics, immunology, and biotechnology [1,2]. There are numerous theoretical and experimental studies devoted to various aspects of this phenomenon, such as the changes of permeability of a cell plasma membrane to various substances in the presence of an electric field [3,4,5,6,7] and the restoration of the state of low permeability after treatment [7,8,9]. When an electric current passes through a cell suspension, it causes heating (Joule heating), and various chemical reactions occur at the surface between the solution and the electrodes (electrolysis). These may include the evolution of gases, the separation of substances, the dissolution of the electrode, or the appearance of new substances in the solution [10,11,12]. When the processes of electrolysis are not taken into consideration, can the results obtained be misinterpreted [13], but it is difficult to improve the experimental procedures
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