Cited by 434Redox biology and oxidative stressCancer, Hypoxia, and MetabolismBiomedical and Chemical Research

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Limitations of oxygen delivery to cells in culture: An underappreciated problem in basic and translational research

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Place, Trenton L., Domann, Frederick E., Case, Adam J.·2017·Free Radical Biology and Medicine

DOI 10.1016/j.freeradbiomed.2017.10.003PMC PMC5699948

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Abstract

Molecular oxygen is one of the most important variables in modern cell culture systems. Fluctuations in its concentration can affect cell growth, differentiation, signaling, and free radical production. In order to maintain culture viability, experimental validity, and reproducibility, it is imperative that oxygen levels be consistently maintained within physiological "normoxic" limits. Use of the term normoxia, however, is not consistent among scientists who experiment in cell culture. It is typically used to describe the atmospheric conditions of a standard incubator, not the true microenvironment to which the cells are exposed. This error may lead to the situation where cells grown in a standard "normoxic" oxygen concentration may actually be experiencing a wide range of conditions ranging from hyperoxia to near-anoxic conditions at the cellular level. This apparent paradox is created by oxygen's sluggish rate of diffusion through aqueous medium, and the generally underappreciated effects that cell density, media volume, and barometric pressure can have on pericellular oxygen concentration in a cell culture system. This review aims to provide an overview of this phenomenon we have termed "consumptive oxygen depletion" (COD), and includes a basic review of the physics, potential consequences, and alternative culture methods currently available to help circumvent this largely unrecognized problem.

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Additional file 5 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
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Additional file 2 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
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Additional file 6 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870327
Additional file 2 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870315
Additional file 4 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870321
Additional file 3 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870318
Additional file 1 of Challenges of aortic valve tissue culture – maintenance of viability and extracellular matrix in the pulsatile dynamic microphysiological system
Dittfeld, C., Winkelkotte, M., Scheer, A., Voigt, E., Schmieder, F., Behrens, S., Jannasch, A., Matschke, K., Sonntag, F., & Tugtekin, S.-M. (2024) · Figshare · doi:10.6084/m9.figshare.26620621
Additional file 4 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
Phelps, J., Hart, D. A., Mitha, A. P., Duncan, N. A., & Sen, A. (2024) · Figshare · doi:10.6084/m9.figshare.26612201
Additional file 7 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870330
Additional file 5 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870324
Additional file 6 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
Phelps, J., Hart, D. A., Mitha, A. P., Duncan, N. A., & Sen, A. (2024) · Figshare · doi:10.6084/m9.figshare.26612207
Additional file 3 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
Phelps, J., Hart, D. A., Mitha, A. P., Duncan, N. A., & Sen, A. (2024) · Figshare · doi:10.6084/m9.figshare.26612198
Additional file 2 of Challenges of aortic valve tissue culture – maintenance of viability and extracellular matrix in the pulsatile dynamic microphysiological system
Dittfeld, C., Winkelkotte, M., Scheer, A., Voigt, E., Schmieder, F., Behrens, S., Jannasch, A., Matschke, K., Sonntag, F., & Tugtekin, S.-M. (2024) · Figshare · doi:10.6084/m9.figshare.26620624
Additional file 8 of In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections
de Buhr, N., Martens, A., Meurer, M., Bonilla, M. C., Söbbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., Kästner, S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.14870333
Additional file 1 of Physiological oxygen conditions enhance the angiogenic properties of extracellular vesicles from human mesenchymal stem cells
Phelps, J., Hart, D. A., Mitha, A. P., Duncan, N. A., & Sen, A. (2024) · Figshare · doi:10.6084/m9.figshare.26612192
Culture conditions shape metabolism and exopolysaccharide production in Piscirickettsia salmonis: implications for vaccine development
Zelada-Cordero, P. A., Altamirano, C., Arancibia‐Díaz, A., Gutiérrez, C. L., Valdez-Cruz, N. A., & Trujillo-Roldán, M. A. (2025) · Aquaculture International · doi:10.1007/s10499-025-02265-5

Authors: Place, T. L., Domann, F. E., & Case, A. J.
Affiliations: University of Iowa (US)
University of Nebraska Medical Center (US)
Corresponding author: Domann, Frederick E.
Case, Adam J.
ORCID: Domann, Frederick E.
Case, Adam J.
Venue: Free Radical Biology and Medicine
Publisher: Elsevier BV
Published: 2017-12
Volume: 113
Pages: 311-322
ISSN: 0891-5849 (ISSN-L)
DOI: 10.1016/j.freeradbiomed.2017.10.003
PMC: PMC5699948
OpenAlex: W2762502468
Cited by: 434
References: 74
Citations by year: 2026: 50 · 2025: 50 · 2024: 68 · 2023: 47 · 2022: 69 · 2021: 72 · 2020: 46 · 2019: 19 · 2018: 12
FWCI: 1.97
Type: journal article
Language: en
Update notices: erratum (2021-01-01)
Open access: hybrid
doi.org
Licence: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor)
Locations: Free Radical Biology and Medicine (publishedVersion, cc-by-nc-nd)
PubMed (publishedVersion)
Europe PMC (PubMed Central) (acceptedVersion)
PubMed Central (acceptedVersion)
Funders: National Heart, Lung, and Blood Institute (R00HL123471)
National Cancer Institute (P30CA086862)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
Health Sciences › Medicine › Physiology (medical) › Biomedical and Chemical Research
Keywords: oxygen delivery, cell culture, normoxia, hyperoxia, anoxia, oxygen diffusion, cell density, atmospheric pressure, culture vessel design, reproducibility
MeSH terms: Cell Count, Cell Differentiation, Cells, Cultured, Diffusion, Humans, Oxygen, Oxygen Consumption, Reproducibility of Results, Cell Hypoxia, Hyperoxia, Cell Culture Techniques, Cell Proliferation, Translational Research, Biomedical

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Limitations of oxygen delivery to cells in culture: An underappreciated problem in basic and translational research

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