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Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies

Nature Protocols volume 5, pages 1540–1551 (2010)Cite this article

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Abstract

Precision-cut tissue slices (PCTS) are viable ex vivo explants of tissue with a reproducible, well defined thickness. They represent a mini-model of the organ under study and contain all cells of the tissue in their natural environment, leaving intercellular and cell-matrix interactions intact, and are therefore highly appropriate for studying multicellular processes. PCTS are mainly used to study the metabolism and toxicity of xenobiotics, but they are suitable for many other purposes. Here we describe the protocols to prepare and incubate rat and human liver and intestinal slices. Slices are prepared from fresh liver by making a cylindrical core using a drill with a hollow bit, from which slices are cut with a specially designed tissue slicer. Intestinal tissue is embedded in cylinders of agarose before slicing. Slices remain viable for 24 h (intestine) and up to 96 h (liver) when incubated in 6- or 12-well plates under 95% O2/5% CO2 atmosphere.

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Figure 1: Setup of the incubation cabinet.
Figure 2: Preparation and incubation of rat or human liver slices.
Figure 3: Preparation and incubation of rat intestinal slices.
Figure 4: Preparation of human intestinal cores.
Figure 5: Histomorphology of slices.

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Acknowledgements

The development of these protocols has been supported by grants from ZonMw, Technology Foundation STW, Organon NV (now MSD); Solvay Pharmaceuticals (now Abbott Healthcare) and Yamanouchi Europe (now Astellas Pharma Inc.). We acknowledge M. van de Bovenkamp, P.M. van Midwoud, A.L. Draaisma, I.H. van Veen-Hof, M.G.L. Elferink, A.A. Khan, J. Plazar, M.G. Rots, D.K.F. Meijer, R.J. Porte, M.J.H. Slooff, K.P. de Jong, V.B. Nieuwenhuijs and R.J. Ploeg for their valuable contributions.

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  1. Peter Olinga, Ruben de Kanter & Esther G van de Kerkhof

    Present address: Present addresses: Division of Pharmaceutical Technology and Biopharmacy, Department of Pharmacy, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands (P.O.), Abbott Healthcare Products BV, Weesp, The Netherlands (R.d.K.), DMPK-Translational Sciences, Novartis Institutes for BioMedical Research, Basel, Switzerland (E.G.v.d.K.).,

Authors and Affiliations

  1. Division of Pharmacokinetics, Department of Pharmacy, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands

    Inge A M de Graaf, Peter Olinga, Marina H de Jager, Marjolijn T Merema, Ruben de Kanter, Esther G van de Kerkhof & Geny M M Groothuis

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  3. Marina H de Jager
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Contributions

All authors contributed extensively to the work presented in this article. P.O., M.T.M., I.A.M.d.G. and G.M.M.G. developed the precision-cut liver slice technique. R.d.K., E.G.v.d.K., I.A.M.d.G., M.H.d.J. and G.M.M.G. developed the precision-cut intestinal slice technique.

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Correspondence to Inge A M de Graaf.

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de Graaf, I., Olinga, P., de Jager, M. et al. Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies. Nat Protoc 5, 1540–1551 (2010). https://doi.org/10.1038/nprot.2010.111

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