Precision-Cut Liver Slices as a New Model to Study Toxicity-Induced Hepatic Stellate Cell Activation in a Physiologic Milieu
Hepatic stellate cell (HSC) activation is a key event in the natural process of wound healing as well as in fibrosis development in liver. Current in vitro models for HSC activation contribute significantly to the understanding of HSC biology and fibrogenesis but still fall far short of recapitulati...
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Published in | Toxicological sciences Vol. 85; no. 1; pp. 632 - 638 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Oxford University Press
01.05.2005
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Subjects | |
Online Access | Get full text |
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Summary: | Hepatic stellate cell (HSC) activation is a key event in the natural process of wound healing as well as in fibrosis development in liver. Current in vitro models for HSC activation contribute significantly to the understanding of HSC biology and fibrogenesis but still fall far short of recapitulating in vivo intercellular functional and anatomic relationships. In addition, when cultured on uncoated plastic, HSC spontaneously activate, which makes HSC activation difficult to regulate or analyze. We have examined whether the use of precision-cut liver slices might overcome these limitations. Liver slices (8 mm diameter, 250 μm thickness) were generated from normal rat liver and incubated for 3 or 16 h with increasing doses of carbon tetrachloride (CCl4). Rat liver slices remained viable during incubation, as shown by minimal enzyme leakage. Expression of markers for HSC activation and the onset of fibrogenesis in the liver slices was studied using real-time PCR and Western blotting. In unstimulated liver slices, mRNA and protein levels of desmin, heat shock protein 47, and αB-crystallin remained constant, indicating quiescence of HSC, whereas Krüppel-like factor 6 expression was increased. In contrast, incubation with CCl4 led to a time- and dose-dependent increase in mRNA expression of all markers and an increased αB-crystallin protein expression. In conclusion, we have developed a technique to induce activation of quiescent HSC in rat liver slices. This model permits the study of toxicity-induced HSC activation within a physiological milieu, not only in animal but ultimately also in human tissue, and could contribute to the reduction of animal experiments. |
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Bibliography: | ark:/67375/HXZ-T3MTN5NW-V 1To whom correspondence should be addressed at University of Groningen, Department of Pharmacokinetics and Drug Delivery, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands. Fax: +31-50–3633247. E-mail: M.van.de.Bovenkamp@rug.nl. local:kfi127 istex:5BB2649641D1E502B863A460EACF78A3A5471255 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 1096-6080 1096-0929 |
DOI: | 10.1093/toxsci/kfi127 |