CoChat
Sign Up
Cited by 152Cancer, Hypoxia, and MetabolismAnimal testing and alternativesAcademic Writing and Publishing

Oxygenation in Cell Culture: Critical Parameters for Reproducibility Are Routinely Not Reported

Open access
Al-Ani, Abdullah, Toms, Derek, Kondro, Douglas, Thundathil, Jarin, Yu, Yang, Ungrin, Mark·2018·PLOS ONE
DOI 10.1371/journal.pone.0204269PMC PMC6191109
Open this paper in CoChat — free account

Abstract

Mammalian cell culture is foundational to biomedical research, and the reproducibility of research findings across the sciences is drawing increasing attention. While many components contribute to reproducibility, the reporting of factors that impact oxygen delivery in the general biomedical literature has the potential for both significant impact, and immediate improvement. The relationship between the oxygen consumption rate of cells and the diffusive delivery of oxygen through the overlying medium layer means parameters such as medium depth and cell type can cause significant differences in oxygenation for cultures nominally maintained under the same conditions. While oxygenation levels are widely understood to significantly impact the phenotype of cultured cells in the abstract, in practise the importance of the above parameters does not appear to be well recognized in the non-specialist research community. On analyzing two hundred articles from high-impact journals we find a large majority missing at least one key piece of information necessary to ensure consistency in replication. We propose that explicitly reporting these values should be a requirement for publication.

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.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0204269

Cites

76 of 76 references verified0 citation errors · 0 not checkable

Statistics

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.

A few free checks a day · papers already verified open instantly · results are public

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 majority of research papers rely on nominal set points and fail to directly verify and report environmental parameters.

    “recent assessments show that the majority of research papers rely on nominal set points and fail to directly verify and report environmental parameters (Al-Ani et al., 2018; Michl et al., 2019; Klein et al., 2021b).”

    From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Al-Ani et al., 2018

    The Results and discussion report that 94% of 200 mammalian-cell-culture papers omitted at least one critical oxygenation variable, and none attempted to calculate oxygen levels around the cells. The Introduction also says incubator settings are often assumed to represent oxygen delivered to cells. This supports the claim for oxygenation in the papers studied, but not its broader wording about research papers and environmental parameters generally; the assessment did not establish how many directly measured oxygen levels. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The lack of monitoring and reporting of environmental conditions in cell culture systems is a pervasive but under-recognized problem.

    “The lack of monitoring and reporting of environmental conditions in cell culture-systems is a pervasive, but under-recognized problem (Hunter, 2017; Al-Ani et al., 2018; Michl et al., 2019; Klein et al., 2021a; Klein et al., 2021b).”

    From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Al-Ani et al., 2018

    The Results and discussion report that 94% of the 200 articles examined omitted at least one critical oxygenation variable, and none attempted to calculate oxygen levels around the cells. The Introduction and Conclusion describe inadequate attention to oxygenation conditions as an under-recognized, prevalent problem. The paper establishes this for oxygenation, not for environmental conditions in cell culture generally. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • Medium changes have been shown to influence oxygen supply and cellular metabolism, as well as synthesis of proteins, DNA, and RNA.

    “Medium changes have been shown to influence oxygen supply and cellular metabolism, as well as synthesis of proteins, DNA, and RNA (Al-Ani et al., 2018; Wright Muelas et al., 2018).”

    From In Vitro research Reproducibility: Keeping up High Standards (2019) · cites this paper as Al-Ani et al., 2018

    The Results and discussion section on key factors affecting oxygen diffusion discusses oxygen delivery after a medium change, and the Introduction section on variations in oxygenation says oxygen availability affects cellular metabolism. Our copy does not say that medium changes affect the synthesis of proteins, DNA, or RNA. Page numbers were not available in the indexed passages. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • 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 Al-Ani et al., 2018

  • Reliable reports of environmental parameters within selected human tissues are presently limited.

    “although reliable reports of environmental parameters within selected human tissues are presently limited (Al-Ani et al., 2018; Ast and Mootha, 2019)”

    From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Al-Ani et al., 2018

    The Results and discussion section (page not provided) finds that oxygenation-related parameters are often missing from reports of mammalian cell cultures. The Introduction discusses oxygen levels in vivo, but does not say that reliable reports of environmental parameters within human tissues are limited. Searches for human-tissue measurements and available in vivo data did not establish that claim. (LLM, conf=0.74)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. [70]Allen CB, Schneider BK, White CW. Limitations to oxygen diffusion and equilibration in in vitro cell exposure systems in hyperoxia and hypoxia. Am J Physiol Cell Mol Physiol. 2001; 281: L1021– L1027. 10.1152/ajplung.2001.281.4.L1021 11557606 · DOI

Cited by

Sorted by citations

Related

Authors
Al-Ani, A., Toms, D., Kondro, D., Thundathil, J., Yu, Y., & Ungrin, M.
Affiliations
  • University of Alberta (CA)
  • University of Calgary (CA)
Corresponding author
  • Al-Ani, Abdullah
  • Toms, Derek
  • Kondro, Douglas
  • Thundathil, Jarin
  • Yu, Yang
  • Ungrin, Mark
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2018-10-16
Volume
13
Issue
10
Pages
e0204269
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0204269
PMC
PMC6191109
OpenAlex
W2896573442
Cited by
152
References
76
Citations by year
2026: 15 · 2025: 20 · 2024: 15 · 2023: 25 · 2022: 29 · 2021: 25 · 2020: 15 · 2019: 8
FWCI
3.63
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
Funders
  • Natural Sciences and Engineering Research Council of Canada (RGPIN-201404874, Graduate Studentship)
  • Institute of Nutrition, Metabolism and Diabetes (MOP-137095)
  • Alberta Diabetes Institute (Graduate Studentship)
  • Alberta Children's Hospital Research Institute (CA) (Graduate Studentship)
  • Alberta Children's Hospital Research Institute (Post-doctoral Fellowship)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
  • Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
  • Social Sciences › Arts and Humanities › History and Philosophy of Science › Academic Writing and Publishing
Keywords
oxygenation, mammalian cell culture, oxygen consumption rate, oxygen diffusion, reproducibility, research reporting practices, publication requirements, culture medium composition, cell phenotype
MeSH terms
Animals, Cells, Cultured, Humans, Mammals, Oxygen, Phenotype, Research Design, Cell Culture Techniques

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