How supraphysiological oxygen levels in standard cell culture affect oxygen-consuming reactions

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Abstract

Most mammalian tissue cells experience oxygen partial pressures in vivo equivalent to 1–6% O 2 (i.e., physioxia). In standard cell culture, however, headspace O 2 levels are usually not actively regulated and under these conditions are ~18%. This drives hyperoxia in cell culture media that can affect a wide variety of cellular activities and may compromise the ability of in vitro models to reproduce in vivo biology. Here, we review and discuss some specific O 2 -consuming organelles and enzymes, including mitochondria, NADPH oxidases, the transplasma membrane redox system, nitric oxide synthases, xanthine oxidase, and monoamine oxidase with respect to their sensitivities to O 2 levels. Many of these produce reactive oxygen and/or nitrogen species (ROS/RNS) as either primary end products or byproducts and are acutely sensitive to O 2 levels in the range from 1% to 18%. Interestingly, many of them are also transcriptional targets of hypoxia-inducible factors (HIFs) and chronic cell growth at physioxia versus 18% O 2 may alter their expression. Aquaporins, which facilitate hydrogen peroxide diffusion into and out of cells, are also regulated by HIFs, indicating that O 2 levels may affect intercellular communication via hydrogen peroxide. The O 2 sensitivities of these important activities emphasize the importance of maintaining physioxia in culture.

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Stuart, J. A., Fonseca, J., Moradi, F., Cunningham, C., Seliman, B., Worsfold, C. R., … Maddalena, L. A. (2018). How supraphysiological oxygen levels in standard cell culture affect oxygen-consuming reactions. Oxidative Medicine and Cellular Longevity. Hindawi Limited. https://doi.org/10.1155/2018/8238459

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