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Primary hepatocyte culture from Oreochromis niloticus fish as a tool for environmental toxicology

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Abstract

Risk assessment studies on aquatic pollution, a threat to water quality and aquatic biota, often rely on animal capture and experimentation. However, ethical concerns surrounding animal use have prompted the development and use of alternative methods. This study investigates the viability of primary hepatocyte cultures from Oreochromis niloticus as an in vitro tool for environmental contaminant screening, focusing on cadmium as a pollutant model. Isolated hepatocytes were cultured on plates using a mixture of supplemented L15/F12 media. Freezing conditions were tested for cell stock preservation. The cultured hepatocytes were exposed to 0.5, 5, and 50 µM of cadmium to assess cell viability, reactive oxygen and nitrogen species, and antioxidant molecules. Results demonstrated a concentration and time-dependent decrease in cell viability upon cadmium exposure. Cadmium exposure also induced an imbalance in the antioxidant defense system, leading to increased reactive oxygen species levels and lipid peroxidation. These findings validate the cadmium toxicity to primary hepatocytes and confirm the applicability of commonly used biomarkers in animal-based ecotoxicology and risk assessment studies. The replication of these biomarkers in primary hepatocyte cultures provides an ethical alternative to reduce animal testing in ecotoxicology. Lastly, the study highlights the potential of primary hepatocyte cultures as an in vitro tool for evaluating the toxicity of environmental contaminants and supports the integration of this tool in future environmental risk assessment studies, promoting sustainable water resource management while reducing reliance on animal experimentation.

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Acknowledgements

The authors thank the Coordination for the Improvement of Higher Education Personnel (CAPES, Ph.D. scholarship—grant n° 88882.382558/2019-01), Council for Scientific and Technological Development (CNPq, Ph.D. scholarship and financial support—grant n° 140931/2018-0), Companhia Paranaense de Energia (COPEL) and Agência Nacional de Energia Elétrica (ANNEL, P&D 06491-0265/2012), Universidade Federal do Paraná (UFPR, equipment and financial support), and the Center for Advanced Fluorescence Technologies (CTAF) at UFPR (technical support).

Funding

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, 88882.382558/2019-01, Jessica Zablocki da Luz, Conselho Nacional de Desenvolvimento Científico e Tecnológico, 140931/2018-0, ALICIANE DE ALMEIDA ROQUE, Agência Nacional de Energia Elétrica, 06491-0265/2012, Ciro Alberto de Oliveira Ribeiro.

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  1. Jessica Zablocki da Luz and Tugstênio Lima de Souza have contributed equally to this study.

Authors and Affiliations

  1. Laboratório de Toxicologia Celular, Departamento de Biologia Celular, Universidade Federal Do Paraná, CEP 81.531-980, Curitiba, PR, Brazil

    J. Zablocki da Luz, T. Lima de Souza, A. de Almeida Roque, M. Mela Prodocimo, C. Alberto de Oliveira Ribeiro & F. Filipak Neto

  2. Copel GeT-SOS/DNGT, CEP 81.200-240, Curitiba, PR, Brazil

    I. Opuskevitch & F. C. Alves da Silva Ferreira

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  1. J. Zablocki da Luz
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Correspondence to J. Zablocki da Luz.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethical approval

The procedures were approved and certified by the Animal Use Ethics Committee of the Biological Sciences Sector of the Federal University of Paraná (CEUA/BIO—UFPR) with certificate number 1286, June 2019.

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Editorial responsibility: Maryam Shabani.

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Zablocki da Luz, J., Lima de Souza, T., de Almeida Roque, A. et al. Primary hepatocyte culture from Oreochromis niloticus fish as a tool for environmental toxicology. Int. J. Environ. Sci. Technol. 21, 2517–2532 (2024). https://doi.org/10.1007/s13762-023-05129-8

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