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Hydrogen peroxide production is affected by oxygen levels in mammalian cell culture.

Abstract：

:Although oxygen levels in the extracellular space of most mammalian tissues are just a few percent, under standard cell culture conditions they are not regulated and are often substantially higher. Some cellular sources of reactive oxygen species, like NADPH oxidase 4, are sensitive to oxygen levels in the range between 'normal' physiological (typically 1-5%) and standard cell culture (up to 18%). Hydrogen peroxide in particular participates in signal transduction pathways via protein redox modifications, so the potential increase in its production under standard cell culture conditions is important to understand. We measured the rates of cellular hydrogen peroxide production in some common cell lines, including C2C12, PC-3, HeLa, SH-SY5Y, MCF-7, and mouse embryonic fibroblasts (MEFs) maintained at 18% or 5% oxygen. In all instances the rate of hydrogen peroxide production by these cells was significantly greater at 18% oxygen than at 5%. The increase in hydrogen peroxide production at higher oxygen levels was either abolished or substantially reduced by treatment with GKT 137831, a selective inhibitor of NADPH oxidase subunits 1 and 4. These data indicate that oxygen levels experienced by cells in culture influence hydrogen peroxide production via NADPH oxidase 1/4, highlighting the importance of regulating oxygen levels in culture near physiological values. However, we measured pericellular oxygen levels adjacent to cell monolayers under a variety of conditions and with different cell lines and found that, particularly when growing at 5% incubator oxygen levels, pericellular oxygen was often lower and variable. Together, these observations indicate the importance, and difficulty, of regulating oxygen levels experienced by cells in culture.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Maddalena LA,Selim SM,Fonseca J,Messner H,McGowan S,Stuart JA

doi

10.1016/j.bbrc.2017.09.037

subject

Has Abstract

pub_date

2017-11-04 00:00:00

pages

246-251

issue

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(17)31797-7

journal_volume

493

pub_type

杂志文章

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Hydrogen peroxide production is affected by oxygen levels in mammalian cell culture.