Toxicity Assessment in 2D vs 3D cultures of Zebrafish liver cells (ZebTox3D)
Ministry of Science, Education and the Youth, Croatian-Slovenian Bilateral Cooperation, 2025-2026
Project Leader:
Leader in Slovenia:
- Matjaž Novak, National Institute of Biology (NIB), Ljubljana, Slovenia
Participants (IMROH):
Participants (NIB):
- Bojana Žegura
- Alja Štern
- Katja Kološa
- Sonja Žabkar
- Iza Rozman
Summary
The zebrafish (Danio rerio) is one of the most important ecotoxicological models for studying the effects of pollutants. In accordance with the 3R principles (Reduce, Refine, Replace), in vitro models utilizing fish cell lines have been developed for xenobiotic toxicity assessment. Zebrafish liver epithelial cells (ZFL) possess the ability to metabolize various compounds and are widely used in toxicological research. However, data obtained from two-dimensional (2D) cultures have shown limitations in predicting in vivo responses, making 3D cultures a more relevant model due to their enhanced cellular differentiation, morphology, and gene expression. While 3D cultures are frequently used in mammalian toxicology studies, their application in ecotoxicology remains scarce. This project aims to establish ZFL 3D cultures as a model for toxicity assessment and to compare the responses of 2D and 3D systems to various pollutants, including bisphenols, nanoparticles, pharmaceutical residues, and polycyclic aromatic hydrocarbons. Cytotoxic and genotoxic effects will be evaluated through assays assessing cell viability, proliferation, migration, induction of cell death, and DNA damage. Comparing results between 2D and 3D models will enable the identification of the most suitable system for predicting in vivo responses. The project builds on a long-standing collaboration between Slovenian and Croatian research groups, leveraging their complementary expertise to advance in vitro methodologies in ecotoxicology. The expected outcomes of this research will contribute to new scientific insights, foster industry collaborations, and facilitate the education of young researchers. The findings will serve as a foundation for future international project applications, including those within the Horizon EU program.
