Limitations of Oxygen Delivery to Cells in Culture: An Underappreciated Problem in Basic and Translational Research
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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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O2 first dissolves at the air-medium interface and then diffuses through the liquid (at least several millimeters) to reach and oxygenate cell microenvironments even as cells undergo exponential growth.
“Specifically, O2 first dissolves at the air-medium interface and then diffuses through the liquid (at least several millimeters) to reach and oxygenate cell microenvironments (Place et al., 2017) even as cells undergo exponential growth.”
From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Place et al., 2017
These processes interact to create a changing environmental gradient from the surface of the medium down to the microenvironment of the cells.
“In concert, these processes interact to create a changing environmental gradient from the surface of the medium down to the microenvironment of the cells (Place et al., 2017).”
From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Place et al., 2017
In the case of O2, medium deoxygenation is caused by the disparity between rates of O2 consumption via cellular metabolism and the replenishment of O2 at the air-medium interface.
“In the case of O2, medium deoxygenation is caused by the disparity between rates of O2 consumption via cellular metabolism and the replenishment of O2 at the air-medium interface (Place et al., 2017; Al-Ani et al., 2018).”
From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Place et al., 2017
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- 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.
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 2017-12
- Volume
- 113
- Pages
- 311-322
- ISSN
- 0891-5849 (ISSN-L)
- 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
- 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 › Surgery › 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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