title: Effect of oxygen concentration and selected protocol factors on viability and gene expression of mouse liver slices.
abstract: Precision cut liver slices (PCLSs) are widely used as a model to study hepatotoxicity. For culturing of PCLS diverse protocols are used which could affect slices viability and results. We aimed to identify the most optimal culture protocol for mouse PCLS. Slices were cultured for 24h under different concentrations of serum, glucose, insulin, and oxygen. Thereafter, slices viability was assessed by biochemical methods. Transcriptome analysis was performed to identify changes introduced by culture at different oxygen concentrations (20%, 40%, 60%, and 80% of oxygen). Medium composition did not affect the slices viability. Although metabolic competence was unaffected by oxygen concentrations, culturing at 80% of oxygen yielded slices with the best biochemical characteristics. The comparison of uncultured vs. cultured slices revealed 2524 genes to be differentially expressed. Genes involved in drug metabolism, peroxisomal and mitochondrial functions were down-regulated while several adaptive/stress response processes were up-regulated. Moreover, 80% of oxygen was the most favorable condition with respect to maintenance of expression of genes involved in drug and energy metabolism. The outcome of this study indicates that mouse PCLS are a valuable tool in research on hepatic functions and toxicity, particularly if they are cultured under a controlled oxygen concentration of 80%.
authors.affiliation: RIKILT - Institute of Food Safety, Wageningen UR, PO Box 230, 6700 AE Wageningen, The Netherlands. ewa.szalowska@wur.nl
authors.forename: Ewa
authors.initials: E
authors.lastname: Szalowska
authors.forename: Geert
authors.initials: G
authors.lastname: Stoopen
authors.forename: Jeroen C W
authors.initials: JC
authors.lastname: Rijk
authors.forename: Si
authors.initials: S
authors.lastname: Wang
authors.forename: Peter J M
authors.initials: PJ
authors.lastname: Hendriksen
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Groot
authors.forename: Jan
authors.initials: J
authors.lastname: Ossenkoppele
authors.forename: Ad A C M
authors.initials: AA
authors.lastname: Peijnenburg
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D020869:Gene Expression Profiling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D010100:Oxygen / Q000494:pharmacology*; D013739:Testosterone / Q000378:metabolism; D046509:Tissue Culture Techniques; D014280:Triglycerides / Q000378:metabolism
chemical_list: D001647:Bile Acids and Salts; D014280:Triglycerides; D013739:Testosterone; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D010100:Oxygen