The push for new alternative toxicity testing platforms has resulted in rapid development of innovative techniques that can be applied to environmental toxicology. This project focuses on the application of these new tools to aquatic toxicology, creating a fish liver microtissue that bridges traditional in vitro techniques and the complexity of whole animal exposures. 3D cultures can provide the benefits of tissue-like cell-cell interactions and the opportunity for longer term cultures, but also present challenges associated with imaging, maintaining viability, and adapting two dimensional assays to 3D systems. Our approach utilizes a commercially available cell line, PLHC-1, to form 3D fish liver microtissues for the assessment of aquatic contaminants. To characterize the fish liver spheroids, histology, differentiation, and response to toxicants were assessed during long term cultures of spheroids. Using benzo(a)pyrene as a model contaminant, we demonstrated the utility of these microtissues for testing polycyclic aromatic hydrocarbons and similar pollutants over several days. Assays for the toxicity of aromatic hydrocarbons and biomarkers of xenobiotic metabolism, were optimized in monolayer and then adapted for use in 3D. By adapting methodologies from tissue pathology and high-throughput in vitro techniques, we integrated morphological assessments and quantification of molecular response for a more complex toxicological assessment. Moving forward, we are utilizing this toxicity testing platform to measure the effects of complex mixtures of carbon nanomaterials and benzo(a)pyrene. This model is applicable for testing real-world contaminant mixtures, and is being developed as a sensitive screening assay for low-level cumulative or chronic exposures. This research is supported by NIEHS Training Grant T32 ES07272, NIEHS Superfund Research Program P42 ES013660, the Institute at Brown for Environment & Society, and the generous support of Donna McGraw Weiss ’89 and Jason Weiss.

Fish liver microtissues for aquatic toxicology: Integrating morphological and molecular responses for in vitro assessment of environmental pollutants

255th National Meeting (2018)

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