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Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity

  • Reviews in Cell Toxicology
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Abstract

A cell line RTL-W1, has been developed from the normal liver of an adult rainbow trout by proteolytic dissociation of liver fragments. RTL-W1 can be grown routinely in the basal medium, L-15, supplemented with 5% fetal bovine serum. In this medium, the cells have been passaged approximately 100 times over an 8-year period. The cells do not form colonies or grow in soft agar. The cultures are heteroploid. The cell shape was predominantly polygonal or epithelial-like, but as cultures became confluent, bipolar or fibroblast-like cells appeared. Among the prominent ultrastructural features of RTL-W1 were distended endoplasmic reticulum and desmosomes. Benzo[a]pyrene was cytotoxic to RTL-W1. Activity for the enzyme, 7-ethoxyresorufin O-deethylase (EROD), which is a measure of the cytochrome P4501A1 protein, increased dramatically in RTL-W1 upon their exposure to increasing concentrations of either β-naphthoflavone (BNF) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). With these properties, RTL-W1 should be useful for studying the expression of the cytochrome P450 enzymes and as a tool for assessing the toxic potency of environmental contaminants.

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Abbreviations

AHH:

aryl hydrocarbon hydroxylase

B[a]P:

benzo[a]pyrene

BNF:

β-naphthoflavone

CHSE-214:

Chinook salmon embryo cells

CYP:

cytochrome P450

DMSO:

dimethyl sulfoxide

ECOD:

7-ethoxycoumarinO-deethylase

EDTA:

ethylenediaminetetraacetic acid

EROD:

7-ethoxyresorufinO-deethylase

FBS:

fetal bovine serum

HEPES:

N-[ω-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid]

H4IIE:

rathepatoma cells

PCB:

polychlorinated biphenyls

PHH:

planar halogenated hydrocarbons

TCDD:

2,3,7,8-tetrachlorodibenzo-p-dioxin

TEM:

transmission electron microscopy

TEFs:

toxic equivalent factors

TS:

0.05 mM Tris 0.2M sucrose

RLE:

rat liver epithelial cells

RTL-W1:

rainbow trout liver-Waterloo 1 cells

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Lee, L.E.J., Clemons, J.H., Bechtel, D.G. et al. Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity. Cell Biol Toxicol 9, 279–294 (1993). https://doi.org/10.1007/BF00755606

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