The cold atmospheric pressure plasma jet interaction with the bio-target is studied in the plasma experiment, 2D fluid model simulations, and with MTT and iCELLigence assays of the viability of cancer cells. It is shown, for the first time, that the use of the grounded substrate under the media with cells considerably amplifies the effect of plasma cancer cell treatment in vitro. Plasma devices with cylindrical and plane geometries generating cold atmospheric plasma jets are developed and tested. The sequence of the streamers which forms the plasma jet is initiated with a voltage of 2.5–6.5 kV applied with the frequency 40 kHz. We suggest using the grounded substrate under the bio-target during the plasma jet treatment of cancer cells. The analysis of the measured plasma spectra and comparison of OH-line intensity for different voltages and gas flow rates allows us to find a range of optimal plasma parameters for the enhanced OH generation. The time-dependent viability is measured for human cell lines, A431 (skin carcinoma), HEK 293 (kidney embryonic cells), and A549 (human lung adenocarcinoma cells) after the plasma jet treatment. The results with cell-based experiments (direct treatment) performed with various plasma jet parameters confirm the maximum efficiency of the treatment with the optimal plasma parameters.

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