Abstract
Cold atmospheric plasma (CAP) is a potential anticancer therapy. CAP has cytotoxic effects when applied either directly to cancer cell cultures or indirectly through plasma-conditioned liquids. This protocol describes how to treat adherent cultures of human cancer cell lines with CAP or plasma-conditioned medium and determine cell viability following treatment. The protocol also includes details on how to quantify the reactive oxygen and nitrogen species present in medium following CAP treatment, using chemical probes using UV-visible or fluorescence spectroscopy. CAP treatment takes ~30 min, and 3 h are required to complete quantification of reactive oxygen and nitrogen species. By providing a standardized protocol for evaluation of the effects of CAP and plasma-conditioned medium, we hope to facilitate the comparison and interpretation of results seen across different laboratories.
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Data availability
Source data are available for Figs. 8–10 at https://upcommons.upc.edu/handle/2117/330395.
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Acknowledgements
This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No. 714793). Authors belong to the SGR2017 1165. M.P.G. and C.C. acknowledge the Generalitat de Catalunya for the ICREA Academia Award. Authors acknowledge the technical support of M. Sánchez.
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J.T. and C.L. contributed to the acquisition, analysis and interpretation of data, optimization of the protocol and writing of the work. F.T. contributed to acquisition and analysis of data, and to revising it. M.P.G. contributed to writing, discussing and revising the work. C.C. conceived the work, contributed to writing and substantively revising it.
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Peer review information Nature Protocols thanks Sander Bekeschus, Hiromasa Tanaka and the other, anonymous reviewer(s) for their contribution to the peer review of this work.
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Key references using this protocol
Tornin, J. et al. Sci. Rep. 9, 10681 (2019): https://doi.org/10.1038/s41598-019-47128-1
Mateu-Sanz, M. et al. Cancers 12, 227 (2020): https://doi.org/10.3390/cancers12010227
Tornin, J. et al. Free Rad. J Biol. Med. 164, 107 (2021): https://doi.org/10.1016/j.freeradbiomed.2020.12.437
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Supplementary Table 1 and Supplementary Methods 1 and 2.
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Tornin, J., Labay, C., Tampieri, F. et al. Evaluation of the effects of cold atmospheric plasma and plasma-treated liquids in cancer cell cultures. Nat Protoc 16, 2826–2850 (2021). https://doi.org/10.1038/s41596-021-00521-5
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DOI: https://doi.org/10.1038/s41596-021-00521-5
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