Design of optimal concentrations for in vitro cytotoxicity experiments
Open accessAbstract
Concentration-dependent cytotoxicity experiments are frequently used in toxicology. Although it has been reported that an adequate choice of concentrations improves the quality of the statistical inference substantially, a recent literature review of three major toxicological journals has shown that the corresponding methods are rarely used in toxicological practice. In this study the performance of different sets of concentrations, also called designs, are analyzed, while the overall goal is to promote the advantages of optimal design procedures and to present a user-friendly guideline for planning new cytotoxicity concentration-response experiments. We compare the frequently used log-equidistant design to a Bayesian design, which is constructed by methods of optimum design theory. Using both a dense data set of concentration-cytotoxicity data of valproic acid (VPA) and regular assay data of 104 substances, the performance of the different designs is analyzed in two scenarios, where detailed previous knowledge on VPA is available or not. The results show that it is critical to apply a specific design strategy to determine optimal concentrations for cytotoxicity testing. In particular, the Bayesian design technique with and without incorporating pre-existing knowledge of a specific test substance resulted in a more precise statistical inference than the other used designs. Finally, we present a guideline for upcoming experiments and an accessible user-friendly Shiny app (see http://shiny.statistik.tu-dortmund.de:8080/app/occe).
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Schürmeyer et al. (2025) illustrated variability of cytotoxicity analyses of valproic acid in six independent experiments (HepG2 cells, CellTiter Blue assay) with 50 concentrations each, demonstrating that even within one cytotoxicity assay framework, concentration-response estimates are uncertain.
“This is supported by data from Schürmeyer et al. (2025) who have illustrated the variability of cytotoxicity analyses of the test compound valproic acid in six independent experiments (HepG2 cells, CellTiter Blue assay) with 50 concentrations each (Fig. 3), demonstrating that even within one cytotoxicity assay framework, concentration-response estimates are uncertain.”
From Cytotoxicity across OECD methods is not a single endpoint: context-dependent roles, variability and implications for decision-making (2026) · cites this paper as Schürmeyer et al., 2025
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“Concentration-dependent cytotoxicity tests are frequently used in toxicology (Vorrink et al. 2018; Brecklinghaus et al. 2022a; Proctor et al 2017; Khetani et al. 2013; Gu et al. 2018).”
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Thoma, T., Löhr, L., Demuth, P., Hambruch, N., Oesch, F., & Landsiedel, R. (2026) · NAM Journal · doi:10.1016/j.namjnl.2026.100126
- Authors
- Schürmeyer, L., Peng, C., Albrecht, W., Brecklinghaus, T., Baur, P., Hengstler, J. G., & Schorning, K.
- Affiliations
- TU Dortmund University (DE)
- Corresponding author
- Schürmeyer, Leonie
- Venue
- Archives of Toxicology
- Published
- 2024-11-16
- Volume
- 99
- Issue
- 1
- Pages
- 357-376
- ISSN
- 0340-5761 (ISSN-L)
- 1432-0738
- PMC
- PMC11748471
- OpenAlex
- W4404407680
- Cited by
- 11
- References
- 1
- Citations by year
- 2026: 6 · 2025: 5
- FWCI
- 2.76
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Archives of Toxicology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Technische Universität Dortmund Eldorado (Technische Universität Dortmund) (submittedVersion, cc-by)
- PUBLISSO (German National Library of Medicine) (submittedVersion, cc-by)
- Funders
- Deutsche Forschungsgemeinschaft (427806116)
- Technische Universität Dortmund
- Topics
- Social Sciences › Decision Sciences › Management Science and Operations Research › Optimal Experimental Design Methods
- Physical Sciences › Mathematics › Statistics and Probability › Statistical Methods in Clinical Trials
- Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
- Keywords
- in vitro cytotoxicity, Bayesian experimental design, valproic acid, prior knowledge incorporation, Shiny app, statistical inference
- MeSH terms
- Bayes Theorem, Cell Survival, Dose-Response Relationship, Drug, Humans, Research Design, Valproic Acid, Toxicity Tests
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