Phase I and phase II metabolic activities are retained in liver slices from mouse, rat, dog, monkey and human after cryopreservation

• Published in: Toxicology in vitro 2004-02, Vol.18 (1), p.121-128
• Main Authors: Martignoni, Marcella, Monshouwer, Mario, de Kanter, Ruben, Pezzetta, Daniele, Moscone, Anna, Grossi, Pietro
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - chemistry; Animals; Biotransformation - physiology; Cryopreservation; Cryopreservation - methods; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - metabolism; Dogs; Drug Evaluation, Preclinical; Formazans - metabolism; Glucuronosyltransferase - biosynthesis; Glucuronosyltransferase - chemistry; Humans; L-Lactate Dehydrogenase - metabolism; L-Lactate Dehydrogenase - secretion; Liver - cytology; Liver - drug effects; Liver - metabolism; Liver slices; Macaca fascicularis; Male; Mice; Rats; Rats, Sprague-Dawley; Species Specificity; Sulfotransferases - biosynthesis; Sulfotransferases - chemistry; Testosterone - metabolism; Testosterone - pharmacology; Tetrazolium Salts - metabolism; Tissue Preservation - methods; Umbelliferones - metabolism; Umbelliferones - pharmacology
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2003.08.005

Precision-cut liver slices are described as a valuable tool for in vitro metabolism studies of potential drug candidates. Recently, some papers reported successful cryopreservation conditions for liver slices, facilitating a broader and more efficient use of the tissue (particularly of human origin). The aim of this study is to evaluate the effect of cryopreservation on both phase I and phase II metabolism in liver slices prepared from mouse, rat, dog, monkey and human, using rapid freezing in the presence of 18% DMSO. Glucuronidation and sulfation activities (phase II) in both freshly prepared and cryopreserved liver slices were determined by rapid LC-MS/MS analyses using 7-hydroxycoumarin as a marker substrate. Testosterone was used as a marker substrate for cytochrome P450 mediated drug metabolism (phase I). Although the metabolic patterns and rates varied among the different species, the phase I and phase II metabolic capacities of the liver slices were well maintained after cryopreservation. Despite the good biotransformation capacity of cryopreserved slices a decrease in viability, expressed as ATP content and LDH leakage, was observed. MTT reduction was well maintained after cryopreservation. The possibility to cryopreserve liver slices will allow a more efficient utilisation of tissue, in particular from human, but also from dog and monkey. Finally, cryopreserved liver slices from mouse, rat, dog, monkey and human with good phase I and II metabolism activities are a useful in vitro tool to compare metabolite profiles of new chemical entities between species.