Establishment of an a priori protocol for the implementation and interpretation of an in-vitro testing battery for the assessment of developmental neurotoxicity
Abstract
In this project we set up a human cell-based DNTin vitro testing strategy that is based on test methods with high readiness and data generated therefrom. The methods underwent afit-for-purpose evaluation that considered four key elements: 1. The test system, 2. the exposure scheme, 3. the assay and analytical endpoint(s) and 4. the classification model. This testing battery was challenged with 119 chemicals for which rich toxicological information was available (for some of them also on their DNT hazard). Testing was performed in 5 test systems measuring 10 DNT-specific endpoints and additional 9 viability/cytotoxicity-related parameters. For approximately half of the compounds, additional and complementary data from DNT in vitro tests was added by the US-EPA. This extended battery was also evaluated. Testing results revealed that the test methods of this current DNT in vitro battery are reliable and reproducible. The endpoints had to a large extent low redundancy. Battery performance, as assessed with compounds well-characterized for DNT hazard had a sensitivity of 82.7% and a specificity of 88.2%. Gap analyses suggested that radial, astro- and microglia as well as myelination endpoints may be added to the battery. Two case studies, one for screening and prioritization of 14 flame retardants, and one on hazard characterization of 2 pesticides, were presented. Hypothetical AOPs were developed based on the latter case study.In conclusion, the DNT testing strategy explored here…
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UKN2/UKN5 assays use murine recombinant Noggin reconstituted in 0.1% BSA.
“UKN2/UKN5 assays use murine recombinant Noggin reconstituted in 0.1% BSA (Masjosthusmann et al., 2020).”
From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Masjosthusmann et al., 2020
Cites
The FAIR Guiding Principles for scientific data management and stewardship
Wilkinson, M. D., Dumontier, M., Aalbersberg, I. J., Appleton, G., Axton, M., Baak, A., Blomberg, N., Boiten, J.-W., da Silva Santos, L. B., Bourne, P. E., et al. (2016) · Scientific Data · doi:10.1038/sdata.2016.18 · Cited by 18,423 · see citing works
Generation of pure GABAergic neurons by transcription factor programming
Yang, N., Chanda, S., Marro, S., Ng, Y.-H., Janas, J. A., Haag, D., Ang, C. E., Tang, Y., Flores, Q., Mall, M., et al. (2017) · Nature Methods · doi:10.1038/nmeth.4291 · Cited by 341 · see citing works
Harrill, J. A., Freudenrich, T., Wallace, K., Ball, K., Shafer, T. J., & Mundy, W. R. (2018) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2018.04.001 · Cited by 86 · see citing works
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Blum, J., Masjosthusmann, S., Bartmann, K., Bendt, F., Dolde, X., Dönmez, A., Förster, N., Holzer, A.-K., Hübenthal, U., Keßel, H. E., et al. (2023) · Chemosphere · doi:10.1016/j.chemosphere.2022.137035 · Cited by 105 · see citing works
Self-citationSachana, M., Shafer, T. J., & Terron, A. (2021) · Biology · doi:10.3390/biology10020086 · Cited by 71 · see citing works
Scientific Validation of Human Neurosphere Assays for Developmental Neurotoxicity Evaluation
Koch, K., Bartmann, K., Hartmann, J., Kapr, J., Klose, J., Kuchovská, E., Pahl, M., Schlüppmann, K., Zühr, E., & Fritsche, E. (2022) · Frontiers in Toxicology · doi:10.3389/ftox.2022.816370 · Cited by 48 · see citing works
Self-citationAuthority), E. (. F. S., Hernandez‐Jerez, A., Coja, T., Paparella, M., Price, A., Henri, J., Focks, A., Louisse, J., Terron, A., Binaglia, M., et al. (2024) · EFSA Journal · doi:10.2903/j.efsa.2024.8759 · Cited by 37 · see citing works
Bridging imaging-based in vitro methods from biomedical research to regulatory toxicology
Piergiovanni, M., Mennecozzi, M., Barale-Thomas, E., Danovi, D., Dunst, S., Egan, D., Fassi, A., Hartley, M., Kainz, P., Koch, K., et al. (2025) · Archives of Toxicology · doi:10.1007/s00204-024-03922-z · Cited by 7 · see citing works
“The integration of test methods identifying glia cell-specific toxicities ( i.e., astrocytes, radial glia, and microglia) and the use of exclusively human-based cell systems shall improve the DNT IVB performance in the future (Masjosthusmann et al. 2020; Serafini et al. 2024).”
Spänle, J. V., Haiber, L. M., & Seeger, B. (2026) · Frontiers in Toxicology · doi:10.3389/ftox.2026.1800157 · Cited by 1 · see citing works
- Authors
- Masjosthusmann, S., Blum, J., Bartmann, K., Dolde, X., Holzer, A., Stürzl, L., Keßel, E. H., Förster, N., Dönmez, A., Klose, J., Pahl, M., Waldmann, T., Bendt, F., Kisitu, J., Suciu, I., Hübenthal, U., Mosig, A., Leist, M., & Fritsche, E.
- Affiliations
- IUF ‐ Leibniz Research Institute for Environmental Medicine, Auf’m Hennekamp 50, 40225 Düsseldorf Germany
- University of Konstanz, UKN – University of Konstanz, Universitätsstr. 10, 78464 Konstanz Germany
- RUB – Ruhr University Bochum, Faculty for Biology and Biotechnolgy, Bioinformatics Group Germany
- Venue
- EFSA Supporting Publications
- Publisher
- Wiley
- Published
- 2020-10
- Volume
- 17
- Issue
- 10
- ISSN
- 2397-8325
- Cited by
- 69
- References
- 3
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