A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio)

Eide, Marta; Rusten, Marte; Male, Rune; Jensen, Knut Helge; Goksøyr, Anders

doi:10.1016/j.aquatox.2013.11.023

Cited by 41 publications

(22 citation statements)

References 68 publications

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“…canaliculatus fed diet with fish oil (Xie et al, ). Together, these results indicate that the expression characteristics of some genes in SCHL cells may be altered compared with in vivo , as reported in many other immortalized hepatocyte lines like ZFL from D. rerio , BRL3A and NRL from rats and HepG2 from humans (Boess et al, ; Harris et al, ; Eide et al, ). Thus, the above results demonstrate that the expression characteristics of key enzyme genes for LC‐PUFA biosynthesis in SCHL cells are similar to those observed in S. canaliculatus liver, which suggests that SCHL can be used as a useful in vitro tool for investigating regulatory mechanisms of LC‐PUFA biosynthesis.…”

Section: Discussionsupporting

confidence: 74%

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Liu

Zhang

Dong

et al. 2017

Journal of Fish Biology

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A hepatocyte line was established from the liver of white-spotted spinefoot Siganus canaliculatus to study the biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFA). The cells from the line, designated S. canaliculatus hepatocyte line (SCHL), grew and multiplied well in Dulbecco's modified Eagle's medium (DMEM)-F12 medium supplemented with 20 mM 4-(2-hydroxyethyl) piperazine-1-ethanesulphonic acid (HEPES), 10% foetal bovine serum (FBS) and 0·5% rainbow trout Oncorhychus mykiss serum at 28° C, showing an epithelial-like morphology and the normal chromosome number of 48 (2n) and have been subcultured for over 60 passages. The identity of the hepatocytes was confirmed by periodic acid Schiff (PAS) staining. The mRNA expression of all genes encoding the key enzymes for LC-PUFA biosynthesis including two desaturases (Δ4 Fad and Δ6-Δ5 Fad) and two elongases (Elovl4 and Elovl5), were detected in all cells from passages 5 to 60 and their expression levels became stable after passage 35 and showed responses to various PUFA incubation. This is similar to the situation determined in the liver of S. canaliculatus that were fed diets containing different fatty acids. These results indicated that SCHL was successfully established and can provide an in vitro tool to investigate lipid metabolism and regulatory mechanisms of LC-PUFA biosynthesis in teleosts, especially marine species.

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Section: Discussionsupporting

confidence: 74%

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Liu

Zhang

Dong

et al. 2017

Journal of Fish Biology

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“…In light of our positive control experiments that showed no remarkable change in mRNA expression of ppar‐α , ‐β and ‐γ , fabp3 or crot in response to the model PPAR‐α agonist WY14643, the ZFL cell system may not be the best choice to address the role of PPAR‐α in the cellular response to PFAS exposure. It has previously been reported that the ZFL cell line does not respond as strongly at the level of gene expression as do primary zebrafish hepatocytes (Eide et al ., ), which may further explain the lack of an obvious effect on transcription of the genes targeted in our study. We also looked for effects on intracellular lipid stores.…”

Section: Discussionmentioning

confidence: 68%

“…The liver has been identified as a major target organ for PFAS contamination in vertebrates (Kennedy et al, 2004;. Therefore, in an effort to study the toxicity and effects of PFOA as well as the shorter chain homologues PFHxA and PFBA on cellular metabolism, we carried out in vitro experiments using zebrafish liver (ZFL) cells, a permanent cell line originating from the Danio rerio ZFL (Eide et al, 2014). In vitro models are cost-effective and allow the testing to be done in a more controlled way to reduce the inter-individual and interexperimental variations that often occur in vivo (Bickley et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Comparativein vitrotoxicity assessment of perfluorinated carboxylic acids

et al. 2016

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Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are synthetic fluorinated compounds that are highly bioaccumulative and persistent organic pollutants. Perfluorooctanoic acid (PFOA), an eight-carbon chain perfluorinated carboxylic acid, was used heavily for the production of fluoropolymers, but concerns have led to its replacement by shorter carbon chain homologues such as perfluorohexanoic acid (PFHxA) and perfluorobutanoic acid (PFBA). However, limited toxicity data exist for these substitutes. We evaluated the toxicity of PFOA, PFHxA and PFBA on a zebrafish liver cell line and investigated the effects of exposure on cell metabolism. Gross toxicity after 96 h of exposure was highest for PFOA and PFO , while PFHxA and PFBA exhibited lower toxicity. Although the structural similarity of these compounds to fatty acids suggests the possibility of interference with the transport and metabolism of lipids, we could not detect any differential expression of peroxisome proliferator-activated receptor (ppar-α, -β and -γ), fabp3 and crot genes after 96 h exposure to up to 10 ppm of the test compounds. However, we observed localized lipid droplet accumulation only in PFBA-exposed cells. To study the effects of these compounds on cell metabolism, we conducted fluorescence lifetime imaging microscopy using naturally fluorescent biomarkers, NADH and FAD. The fluorescence lifetimes of NADH and FAD and the bound/free ratio of each of these coenzymes decreased in a dose- and carbon length-dependent manner, suggesting disruption of cell metabolism. In sum, our study revealed that PFASs with shorter carbon chains are less toxic than PFOA, and that exposure to sublethal dosage of PFOA, PFHxA or PFBA affects cell metabolism. Copyright © 2016 John Wiley & Sons, Ltd.

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“…In the liver of fish fed the low taurine diet, transcript levels of ADO and CSAD were significantly higher than in fish fed the low taurine diet, although no differences were seen in the transcript levels of CDO and TauT [41]. This may reflect the observations of Eide et al who demonstrated a reduced capacity of ZFL cells to upregulate transcription of key genes involved in xenoestrogen responses, compared to primary hepatocytes [55]. Alternatively, it could reflect the differences in the period of exposure or the response of cultured cells that are not under the same metabolic or proliferative control as cells within the animal.…”

Section: Discussionmentioning

confidence: 98%

Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium

et al. 2017

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Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity.

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A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio)

Cited by 41 publications

References 68 publications

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Establishment of a hepatocyte line for studying biosynthesis of long‐chain polyunsaturated fatty acids from a marine teleost, the white‐spotted spinefoot Siganus canaliculatus

Comparativein vitrotoxicity assessment of perfluorinated carboxylic acids

Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium

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