Cell Biology and Toxicology

, Volume 9, Issue 3, pp 279–294

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

  • Lucila E. J. Lee
  • Janine H. Clemons
  • Daniel G. Bechtel
  • Sarah J. Caldwell
  • Kyu-Bo Han
  • Maria Pasitschniak-Arts
  • Dick D. Mosser
  • Niels C. Bols
Reviews in Cell Toxicology

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.

Key Words

cytochrome-P450 7-ethoxyresorufinO-deethylase trout liver cell line 

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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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Lucila E. J. Lee
    • 1
  • Janine H. Clemons
    • 2
  • Daniel G. Bechtel
    • 1
  • Sarah J. Caldwell
    • 1
  • Kyu-Bo Han
    • 1
  • Maria Pasitschniak-Arts
    • 1
  • Dick D. Mosser
    • 2
  • Niels C. Bols
    • 2
  1. 1.Department of Veterinary Anatomy, Western College of Veterinary MedicineUniversity of SaskatchewanSaskatoonCanada
  2. 2.Department of BiologyUniversity of WaterlooWaterlooCanada

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