Oxygenation in cell culture: Critical parameters for reproducibility are routinely not reported.

Abdullah Al-Ani,Douglas Kondro,Mark Ungrin,Derek Toms,Yang Yu,Jarin Thundathil,Zoran Ivanovic

doi:10.1371/journal.pone.0204269

Abdullah Al-Ani, Douglas Kondro + Show 5 more

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https://doi.org/10.1371/journal.pone.0204269

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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.

Highlights

Reproducibility is a critical foundation of science. Shortcomings in this area are a growing concern for preclinical research in particular, given the potential for economic and human health impacts, with some studies reporting more than half of the works they investigated are incompletely reproducible [1,2,3,4,5] (89% [1], 78% [6], 54% [7], and 51% [8])
We propose that journal editors require all manuscripts employing mammalian cell culture to include in the materials and methods a section entitled “Oxygenation considerations”, which would at a minimum, explicitly include cell type, culture chamber specifications, media volume, and cell density both at time of seeding and of experimentation, unless those values are already included elsewhere, in which case their location should be specified in the cover letter to the editor
While any given case of irreproducibility can not necessarily be attributed to a lack of details concerning cell oxygenation status, in order to promote reproducibility in the scientific literature, journal editors should ensure that all published manuscripts employing mammalian tissue culture specify the four critical oxygenation parameters

Summary

Introduction

Reproducibility is a critical foundation of science Shortcomings in this area are a growing concern for preclinical research in particular, given the potential for economic and human health impacts, with some studies reporting more than half of the works they investigated are incompletely reproducible [1,2,3,4,5] (89% [1], 78% [6], 54% [7], and 51% [8]). Several initiatives have been launched to define and mitigate this problem, including “The Reproducibility Project: Cancer Biology”, which aimed to replicate 50 high impact cancer biology articles [10]. This approach has obvious practical and financial limitations, and the flagship project has recently been forced to scale back their objectives from.

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