Analysis of gene expression changes to elucidate the mechanism of chilling injury in precision-cut liver slices

• Published in: Toxicology in vitro 2013-03, Vol.27 (2), p.890-899
• Main Authors: Guan, Na, Blomsma, Sylvia A., Fahy, Gregory M., Groothuis, Geny M.M., de Graaf, Inge A.M.
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Animals; Cell Survival - drug effects; Chilling injury; Cold Temperature - adverse effects; Cryopreservation; Cryoprotectant toxicity; Cryoprotectants; Cryoprotective Agents - adverse effects; Gene Expression Profiling; In Vitro Techniques; Liver - injuries; Liver - metabolism; Male; Micro-array analysis; Oligonucleotide Array Sequence Analysis; Organ Preservation; Rats; Rats, Wistar; RNA - genetics; Tissue banking
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2012.10.009

► Precision-cut liver slices as a model to study the mechanism of chilling injury. ► The mechanism of chilling injury on PCLS was studied by microarray array analysis. ► The mechanism of chilling injury was studied in PLCS without other damaging events. ► Chilling affects stress and immune response and lipid/cholesterol metabolism. The exact mechanism of chilling injury (by a decrease of temperature to sub-physiological values), especially in the intact organ, is yet unknown. Precision-cut liver slices (PCLS), which closely resemble the organ from which they are derived, are an ideal in vitro model to study the mechanism of chilling injury in the intact organ. In the present study we were able to separate chilling injury from other damaging events such as cryoprotectant toxicity and ice-crystal injury and performed micro-array analysis of regulated genes. Pathway analysis revealed that different stress responses, lipid/fatty acid and cholesterol biosynthesis and metabolism were affected by chilling. This indicates that the cell-membrane might be the primary site and sensor for chilling, which may initiate and amplify downstream intracellular signaling events. Most importantly, we were able to identify gene expression responses from stellate cells and Kupffer cells suggesting the involvement of all liver cell types in the injury. In conclusion, a broad spectrum of previously unknown gene expression changes induced by chilling was identified in the tissue. This is the first report of a systematic investigation on the mechanism of chilling injury in integrated tissue by micro-array analysis under conditions in which other sources of injury are minimal.