The differential cytotoxicity of methotrexate in rat hepatocyte monolayer and spheroid cultures

It is important to assess the usefulness of long-term in vitro liver models for studying chronic toxicity, since acute assays may not reflect the in vivo situation. A potential long-term hepatocyte culture (i.e. liver spheroids) was investigated and compared to primary rat hepatocyte monolayer cultu...

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Bibliographic Details
Published in:Toxicology in vitro 2000-10, Vol.14 (5), p.475-485
Main Authors: Walker, T.M, Rhodes, P.C, Westmoreland, C
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - metabolism
Alanine Transaminase - metabolism
Animals
Aspartate Aminotransferases - metabolism
Biological and medical sciences
chronic toxicity
Dose-Response Relationship, Drug
Drug toxicity and drugs side effects treatment
Glutathione - metabolism
L-Lactate Dehydrogenase - metabolism
Liver - drug effects
Liver - pathology
long-term liver culture
Male
Medical sciences
methotrexate
Methotrexate - toxicity
Pharmacology. Drug treatments
Rats
Rats, Wistar
spheroids
Spheroids, Cellular - drug effects
Spheroids, Cellular - enzymology
Spheroids, Cellular - pathology
Toxicity Tests
Toxicity: digestive system
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Summary:It is important to assess the usefulness of long-term in vitro liver models for studying chronic toxicity, since acute assays may not reflect the in vivo situation. A potential long-term hepatocyte culture (i.e. liver spheroids) was investigated and compared to primary rat hepatocyte monolayer cultures following exposure to methotrexate (MTX), a well-documented chronic hepatotoxin. Following up to 7 days' treatment with MTX, cultures were morphologically assessed and assayed for enzyme leakage, intracellular reduced glutathione (GSH) and adenosine triphosphate (ATP). Spheroids maintained higher concentrations of GSH over the 14-day culture and ATP was maintained, but at a concentration not significantly different from monolayer cultures. Treatment of monolayer cultures resulted in concentration-related decreases in GSH and ATP, accompanied by enzyme leakage. In contrast, only ATP was affected following treatment of spheroids for 7 days. Spheroids appeared to be less sensitive to exposure to MTX, when compared with monolayer cultures. This may result from the maintenance of cellular functions, or from the lack of compound penetration into the three-dimensional spheroid structure. Therefore, the usefulness of spheroids to chronic in vitro toxicity testing may be limited.
ISSN:0887-2333
1879-3177
DOI:10.1016/S0887-2333(00)00036-9