4.1 Article

Organotypic liver culture in a fluid-air interface using slices of neonatal rat and adult human tissue - a model of fibrosis in vitro

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Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/S1056-8719(03)00042-X

Keywords

hepatic stellate cells; human; liver fibrosis; mouse; organotypic method; rat; slice culture

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Introduction: Fibrosis is the common end stage of most liver disease but there is no effective treatment currently available. We hypothesised that if viability of liver tissue slice culture could be improved, it should be possible to develop a model of liver fibrosis in vitro that could advance the development of antifibrotic therapy while at the same time reducing the need to use in vivo models. We have adapted a slice culture technique developed originally for organotypic culture of neural tissue to the liver. Methods: slices of neonatal rat or adult human liver, 100-400-mum thick, were cut and cultured on nitrocellulose inserts at the air/fluid interface for up to 28 days. Results: Hepatocytes expressed albumin by immunocytochemistry for up to 10 days and were viable for up to 21 days during which time new structures appeared, including cytokeratin 19 positive bile ductular structures and bands of smooth muscle actin positive stellate cells associated with new reticulin positive matrix. Smooth muscle actin expression by stellate cells could be pharmacologically inhibited by SDZ-RAD (everolimus). Discussion: In conclusion, we have successfully developed a novel model of liver culture, which may prove useful in both studies of the mechanisms of liver fibrosis and in developing therapeutic strategies. (C) 2003 Elsevier Inc. All rights reserved.

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