Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies.
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% O(2)/5% CO(2) atmosphere.
de Graaf
Inge A M
IA
Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands. i.a.m.de.graaf@rug.nl
Olinga
Peter
P
de Jager
Marina H
MH
Merema
Marjolijn T
MT
de Kanter
Ruben
R
van de Kerkhof
Esther G
EG
Groothuis
Geny M M
GM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects / Q000473:pathology; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008867:Microtomy / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D046509:Tissue Culture Techniques / Q000295:instrumentation; D015262:Xenobiotics / Q000378:metabolism* / Q000633:toxicity
D015262:Xenobiotics; D000255:Adenosine Triphosphate