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  • Precision-cut liver slices of Atlantic cod (Gadus morhua): an in vitro system for studying the effects of environmental contaminants.

Precision-cut liver slices of Atlantic cod (Gadus morhua): an in vitro system for studying the effects of environmental contaminants.

Aquatic toxicology (Amsterdam, Netherlands) (2013-11-26)
M Eide, O A Karlsen, H Kryvi, P A Olsvik, A Goksøyr
ABSTRACT

The Atlantic cod (Gadus morhua) is an economically important species commonly consumed by humans. The widespread distribution of cod in the North Atlantic Ocean makes it vulnerable to effluents from human activities, such as coastal industries and offshore petroleum exploration. It has been demonstrated that many effluents have adverse effects on cod reproduction and health, e.g. by disrupting endocrine signaling pathways. The liver, expressing important components of the biotransformation and the endocrine system, is one of the main target organs. Thus, reliable and reproducible in vitro systems of the liver are important for studying effects of environmental contaminants. The aim of this study was to investigate precision-cut liver slices (PCLS) as an alternative in vitro system for toxicological studies of the Atlantic cod liver. Slices of 8 mm in diameter and 250 μm thickness were prepared and cultivated from immature cod. Several analyses to measure the liver slice viability were performed: enzyme assays, histology, and morphometric analysis, all confirming cell viability for up to 72 h in culture. The liver slices were also exposed to two well-known model environmental contaminants, β-naphthoflavone (BNF) and 17α-ethynylestradiol (EE2), representing established agonists for the aryl hydrocarbon receptor (AHR) and the estrogen receptor (ER), respectively. The results showed increased transcription of the target genes cytochrome P450 1A (CYP1A) and vitellogenin (VTG), both well-established biomarkers for exposure of fish to the selected compounds. In conclusion, PCLS is a promising in vitro system for toxicological studies of cod liver cells. The liver slices are viable in culture for several days and respond to environmental contaminants in a dose- and time-specific manner.

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Sigma-Aldrich
17α-Ethynylestradiol, ≥98%
Supelco
Ethinyl Estradiol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
β-Naphthoflavone, ≥98%
USP
Ethinyl estradiol, United States Pharmacopeia (USP) Reference Standard
Ethinylestradiol, European Pharmacopoeia (EP) Reference Standard
Supelco
17α-Ethynylestradiol solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
Ethinylestradiol for system suitability, European Pharmacopoeia (EP) Reference Standard