Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity.
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 beta-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.
Lee
L E
LE
Department of Veterinary Anatomy, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
Clemons
J H
JH
Bechtel
D G
DG
Caldwell
S J
SJ
Han
K B
KB
Pasitschniak-Arts
M
M
Mosser
D D
DD
Bols
N C
NC
D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity; D001571:Benzoflavones / Q000633:toxicity; D002460:Cell Line; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004784:Environmental Monitoring; D004787:Environmental Pollution; D004790:Enzyme Induction / Q000187:drug effects; D007621:Karyotyping; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D008854:Microscopy, Electron; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000378:metabolism*; D010088:Oxidoreductases / Q000096:biosynthesis; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity; D019324:beta-Naphthoflavone
D001571:Benzoflavones; D000072317:Polychlorinated Dibenzodioxins; D001564:Benzo(a)pyrene; D019324:beta-Naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D010105:Oxygenases; D019363:Cytochrome P-450 CYP1A1