What is it about?

In current study, we focused on methodology of high-impact cell culture papers to evaluate adequacy of the evidence they provided to support their claims. We identified common claims put forward (such as changes in viability, cytotoxicity, proliferation/growth rate and cell death) and investigated if the experimental design is appropriate to measure related claims. We detected multiple cases of misinterpretation of assay results, inappropriate (or completely lacking) evidence for claims, missing information and other issues.

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Why is it important?

There is an alarming quality crisis in cell culture studies. Confusion regarding the terminology and misinterpretation of assays are amongst the causes. Here we report that even high-impact studies fail to justifiably interpret cell culture methods. We also provide clear and distinct definitions for each term (viability, cytotoxicity, proliferation/growth rate and cell death) and discuss best ways to measure them.


I made an observation on MTT (or similar reduction) assays during my masters and PhD. Those assays were essentially measuring cellular metabolic activity (which is hardly a proper evidence for viability) but were utilized by researchers as a way to measure viability, cytotoxicity, proliferation rate and even cell death, without any change in experimental design. This initial observation led to an investigation for a nomenclature and a guide on cell culture practices, which was failed as there were none. The confusion was everywhere from ResearchGate questions by PhD students to papers published in high-impact journals. To provide a clarity and investigate the situation more systematically we wrote down clear and distinctive definitions for each term and then discussed (and decided on) best ways to measure them. We then investigated high-impact publications on cell culture to evaluate the methodology. Maybe not so surprisingly we detected numerous issues regarding the misinterpretation of assays. We published our findings in PLOS ONE. We hope that this study can raise some awareness on this topic and can be used as a mini-guide for cell culture practices.

PhD Ali Burak Özkaya
Izmir Ekonomi Universitesi

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This page is a summary of: From viability to cell death: Claims with insufficient evidence in high-impact cell culture studies, PLoS ONE, February 2022, PLOS, DOI: 10.1371/journal.pone.0250754.
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