Role of peroxide in AC electrical field exposure effects on Friend murine erythroleukemia cells during dielectrophoretic manipulations

摘要

The effects of AC field exposure on the viability and proliferation of mammalian cells under conditions appropriate for their dielectrophoretic manipulation and sorting were investigated using DS19 murine erythroleukemia cells as a model system. The frequency range 100 Hz-10 MHz and medium conductivities of 10 mS/m, 30 mS/m and 56 mS/m were studied for fields generated by applying signals of up to 7V peak to peak (p-p) to a parallel electrode array having equal electrode widths and gaps of 100 μm. Between 1 kHz and 10 MHz, cell viability after up to 40 min of field exposure was found to be above 95% and cells were able to proliferate. However, cell growth lag phase was extended with decreasing field frequency and with increasing voltage, medium conductivity and exposure duration. Modified growth behavior was not passed on to the next cell passage, indicating that field exposure did not cause permanent alterations in cell proliferation characteristics. Cell membrane potentials induced by field exposure were calculated and shown to be well below values typically associated with cell damage. Furthermore, medium treated by field exposure and then added to untreated cells produced the same modifications of growth as exposing cells directly, and these modifications occurred only when the electrode polarization voltage exceeded a threshold of ~0.4 V p-p. These findings suggested that electrochemical products generated during field exposure were responsible for the changes in cell growth. Finally, it was found that hydrogen peroxide was produced when sugar-containing media were exposed to fields and that normal cell growth could be restored by addition of catalase to the medium, whether or not field exposure occurred in the presence of cells. These results show that AC fields typically used for dielectrophoretic manipulation and sorting of cells do not damage DS19 cells and that cell alterations arising from electrochemical effects can be completely mitigated.