The DOI matches its Crossref record; 74 of the 74 references it cites were found in a registry or an index.
Limitations of oxygen delivery to cells in culture: An underappreciated problem in basic and translational research
Open accessAbstract
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.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The cited source says what the claim attributes to it.
In the citing paper
“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
The cited source says what the claim attributes to it.
In the citing paper
“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
The cited source says what the claim attributes to it.
In the citing paper
“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
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
The Ki-67 protein: From the known and the unknown
Scholzen, T. E., & Gerdes, J. (2000) · Journal of Cellular Physiology · doi:10.1002/(sici)1097-4652(200003)182:3 · Cited by 4,707
Kim, J.-W., Tchernyshyov, I., Semenza, G. L., & Dang, C. V. (2006) · Cell Metabolism · doi:10.1016/j.cmet.2006.02.002 · Cited by 3,905
HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption
Papandreou, I., Cairns, R. A., Fontana, L., Lim, A. L., & Denko, N. C. (2006) · Cell Metabolism · doi:10.1016/j.cmet.2006.01.012 · Cited by 2,267
THE THEORY AND APPLICATIONS OF THE EXCHANGE OF INERT GAS AT THE LUNGS AND TISSUES
Kety, S. S. (1951) · Pharmacological Reviews · doi:10.1016/s0031-6997(25)07246-1 · Cited by 1,665
Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices
Halldorsson, S., Lucumi, E., Gómez-Sjöberg, R., & Fleming, R. M. T. (2015) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2014.07.029 · Cited by 1,027
(1943) · JNCI Journal of the National Cancer Institute · doi:10.1093/jnci/4.2.165 · Cited by 833
Characterization of the human prolyl 4-hydroxylases that modify the hypoxia-inducible factor
Hirsilä, M., Koivunen, P., Günzler, V., Kivirikko, K. I., & Myllyharju, J. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m304982200 · Cited by 794
Krogh, A. (1919) · The Journal of Physiology · doi:10.1113/jphysiol.1919.sp001838 · Cited by 633
The Multiple Origins of Complex Multicellularity
Knoll, A. H. (2011) · Annual Review of Earth and Planetary Sciences · doi:10.1146/annurev.earth.031208.100209 · Cited by 562
The supply of oxygen to the tissues and the regulation of the capillary circulation
Krogh, A. (1919) · The Journal of Physiology · doi:10.1113/jphysiol.1919.sp001844 · Cited by 530
Benita, Y., Kikuchi, H., Smith, A. D., Zhang, M. Q., Chung, D. C., & Xavier, R. J. (2009) · Nucleic Acids Research · doi:10.1093/nar/gkp425 · Cited by 485
Wittenberg, B. A., & Wittenberg, J. B. (1989) · Annual Review of Physiology · doi:10.1146/annurev.ph.51.030189.004233 · Cited by 469
Biological detection by optical oxygen sensing
Papkovsky, D. B., & Dmitriev, R. I. (2013) · Chemical Society Reviews · doi:10.1039/c3cs60131e · Cited by 450
Oxygen gradients correlate with cell density and cell viability in engineered cardiac tissue
Radisic, M., Malda, J., Epping, E., Geng, W., Langer, R., & Vunjak‐Novakovic, G. (2006) · Biotechnology and Bioengineering · doi:10.1002/bit.20722 · Cited by 399
Pedraza, E., Coronel, M. M., Fraker, C. A., Ricordi, C., & Stabler, C. L. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1113560109 · Cited by 399
The rate of oxygen utilization by cells
Wagner, B. A., Venkataraman, S., & Buettner, G. R. (2011) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2011.05.024 · Cited by 380
Davis, W. B., Rennard, S. I., Bitterman, P. B., & Crystal, R. G. (1983) · New England Journal of Medicine · doi:10.1056/nejm198310133091502 · Cited by 343
Finch, C. A., & Lenfant, C. J. M. (1972) · New England Journal of Medicine · doi:10.1056/nejm197202242860806 · Cited by 320
The Redox Basis of Epigenetic Modifications: From Mechanisms to Functional Consequences
Cyr, A. R., & Domann, F. E. (2010) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3492 · Cited by 301
Self-citationEnergetics of interconversion reactions of oxyradicals
Koppenol, W. H., & Butler, J. (1985) · Advances in free radical biology and medicine · doi:10.1016/8755-9668(85)90005-5 · Cited by 295
Increased Prolyl 4-Hydroxylase Domain Proteins Compensate for Decreased Oxygen Levels
Stiehl, D. P., Wirthner, R., Köditz, J., Spielmann, P., Camenisch, G., & Wenger, R. H. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m601719200 · Cited by 286
Regulation of Tissue Oxygenation
Pittman, R. N. (2011) · Colloquium Series on Integrated Systems Physiology From Molecule to Function · doi:10.4199/c00029ed1v01y201103isp017 · Cited by 258
Ehrismann, D., Flashman, E., Genn, D. N., Mathioudakis, N., Hewitson, K. S., Ratcliffe, P. J., & Schofield, C. J. (2007) · Biochemical Journal · doi:10.1042/bj20061151 · Cited by 225
Maina, J. N., & West, J. B. (2005) · Physiological Reviews · doi:10.1152/physrev.00022.2004 · Cited by 211
Fluid mechanics of a spinner‐flask bioreactor
Sucosky, P., Osorio, D., Brown, J. B., & Neitzel, G. P. (2003) · Biotechnology and Bioengineering · doi:10.1002/bit.10788 · Cited by 208
Hyperoxia-Induced Reactive Oxygen Species Formation in Pulmonary Capillary Endothelial Cells In Situ
Brueckl, C., Kaestle, S. M., Kerem, A., Habazettl, H., Krombach, F., Kuppe, H., & Kuebler, W. M. (2005) · American Journal of Respiratory Cell and Molecular Biology · doi:10.1165/rcmb.2005-0223oc · Cited by 207
Rapid reprogramming of epigenetic and transcriptional profiles in mammalian culture systems
Nestor, C. E., Ottaviano, R., Reinhardt, D., Cruickshanks, H. A., Mjoseng, H. K., McPherson, R. C., Lentini, A., Thomson, J. P., Dunican, D. S., Pennings, S., et al. (2015) · Genome biology · doi:10.1186/s13059-014-0576-y · Cited by 178
Cooper, C. E., & Giulivi, C. (2007) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00310.2006 · Cited by 176
Herst, P. M., & Berridge, M. V. (2006) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/j.bbabio.2006.11.018 · Cited by 171
Pericellular PO2 and O2 Consumption in Monolayer Cell Cultures
Metzen, E., Wolff, M., Fandrey, J., & Jelkmann, W. (1995) · Respiration Physiology · doi:10.1016/0034-5687(94)00125-j · Cited by 153
TET1 regulates hypoxia-induced epithelial-mesenchymal transition by acting as a co-activator
Tsai, Y.-P., Chen, H.-F., Chen, S.-Y., Cheng, W.-C., Wang, H.-W., Shen, Z.-J., Song, C., Teng, S.-C., He, C., & Wu, K.-J. (2014) · Genome biology · doi:10.1186/s13059-014-0513-0 · Cited by 152
Allen, C. B., Schneider, B. K., & White, C. W. (2001) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.2001.281.4.l1021 · Cited by 136
Sheta, E. A., Trout, H., Gildea, J. J., Harding, M. A., & Theodorescu, D. (2001) · Oncogene · doi:10.1038/sj.onc.1204972 · Cited by 133
(1987) · Environmental Pollution · doi:10.1016/0269-7491(87)90191-6 · Cited by 131
Koivunen, P., Hirsilä, M., Kivirikko, K. I., & Myllyharju, J. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m604628200 · Cited by 129
Cell and tissue culture : laboratory procedures in biotechnology
Doyle, A., & Griffiths, J. B. (1998) · Wiley eBooks · Cited by 128
Nitric oxide regulation of mitochondrial oxygen consumption I: cellular physiology
Giulivi, C., Kato, K., & Cooper, C. E. (2006) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00307.2006 · Cited by 122
Cellular energy metabolism during hypoxia
Gutiérrez, G. (1991) · Critical Care Medicine · doi:10.1097/00003246-199105000-00008 · Cited by 120
Oxygen and glucose deprivation induces widespread alterations in mRNA translation within 20 minutes
Andreev, D. E., O’Connor, P. B. F., Zhdanov, A. V., Dmitriev, R. I., Shatsky, I. N., Papkovsky, D. B., & Baranov, P. V. (2015) · Genome biology · doi:10.1186/s13059-015-0651-z · Cited by 111
Monitoring pH and dissolved oxygen in mammalian cell culture using optical sensors
Naciri, M., Kuystermans, D., & Al-Rubeai, M. (2008) · Cytotechnology · doi:10.1007/s10616-008-9160-1 · Cited by 111
Pericellular oxygen depletion during ordinary tissue culturing, measured with oxygen microsensors
Pettersen, E. O., Larsen, L. H., Ramsing, N. B., & Ebbesen, P. (2005) · Cell Proliferation · doi:10.1111/j.1365-2184.2005.00345.x · Cited by 110
Gstraunthaler, G. J. A., Seppi, T., & Pfaller, W. (1999) · Cellular Physiology and Biochemistry · doi:10.1159/000016312 · Cited by 109
Abaci, H. E., Truitt, R., Luong, E., Drazer, G., & Gerecht, S. (2010) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00484.2009 · Cited by 108
Dynamics of muscle microcirculatory and blood–myocyte O2flux during contractions
Poole, D. C., Copp, S. W., Hirai, D. M., & Musch, T. I. (2010) · Acta Physiologica · doi:10.1111/j.1748-1716.2010.02246.x · Cited by 106
TET enzymes and DNA hydroxymethylation in neural development and function — How critical are they?
Santiago, M., Antunes, C., Guedes, M., Sousa, N., & Marques, C. J. (2014) · Genomics · doi:10.1016/j.ygeno.2014.08.018 · Cited by 102
Dissolved oxygen concentration in culture medium: assumptions and pitfalls
Newby, D., Marks, L., & Lyall, F. (2004) · Placenta · doi:10.1016/j.placenta.2004.07.002 · Cited by 89
Tissue engineering methods and protocols
Morgan, J. R., & Yarmush, M. L. (1999) · Cited by 88
Kinetics of gas diffusion in mammalian cell culture systems. II. Theory
McLimans, W. F., Blumenson, L. E., & Tunnah, K. V. (1968) · Biotechnology and Bioengineering · doi:10.1002/bit.260100604 · Cited by 88
Chapple, S. J., Keeley, T. P., Mastronicola, D., Arno, M. J., Vizcay‐Barrena, G., Fleck, R. A., Siow, R. C. M., & Mann, G. E. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.12.013 · Cited by 87
Cell Respiration Under Hypoxia: Facts and Artefacts in Mitochondrial Oxygen Kinetics
Scandurra, F. M., & Gnaiger, E. (2009) · Advances in experimental medicine and biology · doi:10.1007/978-1-4419-1241-1_2 · Cited by 86
An Improved Technic for Massive Tissue Culture
Gey, G. O. (1933) · American Journal of Cancer · doi:10.1158/ajc.1933.752 · Cited by 84
Diffusion of Oxygen in Plasma and Blood
Goldstick, T. K., Ciuryla, V. T., & Zuckerman, L. (1976) · Advances in experimental medicine and biology · doi:10.1007/978-1-4684-3273-2_23 · Cited by 76
Oxygen microenvironment and respiratory oscillations in cultured mammalian cells
Werrlein, R. J., & Glinos, A. D. (1974) · Nature · doi:10.1038/251317a0 · Cited by 73
Use of a New Buffer in the Culture of Animal Cells
Williamson, J. D., & Cox, P. (1968) · Journal of General Virology · doi:10.1099/0022-1317-2-2-309 · Cited by 72
Ali, S. S., Marcondes, M.-C. G., Bajova, H., Dugan, L. L., & Conti, B. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.155432 · Cited by 71
Wolff, M., Fandrey, J., & Jelkmann, W. (1993) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.1993.265.5.c1266 · Cited by 70
A mechanistic overview of TET-mediated 5-methylcytosine oxidation
Ponnaluri, V. K. C., Maciejewski, J. P., & Mukherji, M. (2013) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2013.05.077 · Cited by 55
Unforeseen decreases in dissolved oxygen levels affect tube formation kinetics in collagen gels
Abaci, H. E., Truitt, R., Tan, S., & Gerecht, S. (2011) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00074.2011 · Cited by 50
Kinetics of gas diffusion in mammalian cell culture systems. I. Experimental
McLimans, W. F., Crouse, E. J., Tunnah, K. V., & Moore, G. E. (1968) · Biotechnology and Bioengineering · doi:10.1002/bit.260100603 · Cited by 46
Prolyl-hydroxylase 3: evolving roles for an ancient signaling protein
Domann, F. E., & Place (2013) · Hypoxia · doi:10.2147/hp.s50091 · Cited by 45
Self-citationIn vitro intermittent hypoxia: challenges for creating hypoxia in cell culture
Baumgardner, J. E., & Otto, C. M. (2003) · Respiratory Physiology & Neurobiology · doi:10.1016/s1569-9048(03)00077-6 · Cited by 44
Xiao, W., Perry, G., Komori, K., & Sakai, Y. (2015) · Integrative Biology · doi:10.1039/c5ib00170f · Cited by 38
Significance of Flocculation Tests
Maclagan, N. F. (1948) · BMJ · doi:10.1136/bmj.2.4585.892 · Cited by 36
Papkovsky, D. B., & Zhdanov, A. V. (2016) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2016.09.018 · Cited by 34
Pericellular oxygen concentration of cultured primary human trophoblasts
Chen, B., Longtine, M. S., & Nelson, D. M. (2012) · Placenta · doi:10.1016/j.placenta.2012.11.011 · Cited by 22
Klatt, S., Stangl, H., Kunath, J., Lowin, T., Pongratz, G., & Straub, R. H. (2016) · Brain Behavior and Immunity · doi:10.1016/j.bbi.2016.02.006 · Cited by 16
Davis, W. B., Rennard, S. I., Bitterman, P. B., & Crystal, R. G. (1984) · Survey of Anesthesiology · doi:10.1097/00132586-198408000-00003 · Cited by 15
Rigual-Gonzalez, Y., Gómez, L., Núñez, J., Vergara, M., Díaz‐Barrera, A., Berríos, J., & Altamirano, C. (2016) · Chemical Engineering Science · doi:10.1016/j.ces.2016.06.051 · Cited by 8
Biomedical advances from tissue culture
Okamoto, T., Sato, J., Barnes, D. W., & SATO, G. H. (2013) · Cytotechnology · doi:10.1007/s10616-013-9591-1 · Cited by 8
Kino‐oka, M., Kagita, S., Nadzir, M. M., Inoue, H., Sugawara, K., & Taya, M. (2010) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2010.03.009 · Cited by 7
Morgan, J. R., & Yarmush, M. L. (1998) · Humana Press eBooks · doi:10.1385/0896035166 · Cited by 6
Los Alamos Climatology 2016 Update
Los Alamos National Lab. (LANL), L. A., Bruggeman, D., & USDOE (2017) · doi:10.2172/1343690 · Cited by 6
Oxygen Monitoring in Cell Cultures
Lynn, S. G. (2011) · Genetic Engineering & Biotechnology News · doi:10.1089/gen.31.6.20 · Cited by 3
Cited by
Sorted by citations
Defining roles of specific reactive oxygen species (ROS) in cell biology and physiology
Sies, H., Belousov, V. V., Chandel, N. S., Davies, M. J., Jones, D. P., Mann, G. E., Murphy, M. P., Yamamoto, M., & Winterbourn, C. (2022) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-022-00456-z · Cited by 1,735
Keeley, T. P., & Mann, G. E. (2019) · Physiological Reviews · doi:10.1152/physrev.00041.2017 · Cited by 329
Cell Clustering Promotes a Metabolic Switch that Supports Metastatic Colonization
Labuschagne, C. F., Cheung, E. C., Blagih, J., Domart, M., & Vousden, K. H. (2019) · Cell Metabolism · doi:10.1016/j.cmet.2019.07.014 · Cited by 225
Neutrophil intrinsic and extrinsic regulation of NETosis in health and disease
Poli, V., & Zanoni, I. (2022) · Trends in Microbiology · doi:10.1016/j.tim.2022.10.002 · Cited by 223
Impact of Culture Medium on Cellular Interactions in in vitro Co-culture Systems
Vis, M. A. M., Ito, K., & Hofmann, S. (2020) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.00911 · Cited by 210
Metadata
- 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 › 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
