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Livrables
The in vitro model of precision-cut tissue slice cultures may be used to study and to analyse the capacity of chemicals to induce organ-specific pharmaco-toxicological effects. In addition, the successful cold-and cryo-preservation of human liver and kidneys promotes an efficient utilisation of available human tissues, it will result in a reduction in the use of animal for experimentation, and will enhance the use of in vitro alternative methods. Actually, our results show clearly that tissue slice cultures allow an integrative approach by combining procedures of cell biology, biochemistry, molecular biology, histology and histochemistry. The maintenance of metabolic and differentiated functions in long-term incubations give the oportunity to study the influence of cell-cell interactions (for instance non-parenchymal cells in liver tissue), the in vitro induction of phase I and phase II enzymes and xenobiotic metabolism as well as xenobiotic-induced toxicity.
-We determined the optimal conditions to prepare and to culture fresh rat liver, kidney and lung slices. We established the procedures to cold- and cryo-preserved rat and human tissue slices. We showed the maintenance of cell differentiated functions and cell-cell interactions for long-term incubation times (24-72 hours). Finally, both phase I and phase II metabolising enzymes may be induced in vitro.
-This in vitro system can be used in pharmacotoxicological screening or in fundamental research by taking other tissues and/or to study the influence of environmental factors, nutritional status, etc.
-Cell lines have been only used in the field of in vitro toxicology. The main disadvantage of these systems (primary cultures, cells in suspension, immortalized cell lines, co-cultures, etc) is the loss of their tissue architecture making difficult the study of cell-cell interactions and a concomitant histological and biochemical approach.
-For that reason, the experimental data that we obtained are fairly close to the in vivo conditions. In addition, by using cryopreserved tissue material (both humans and rats), we can best assessed the drawbacks and pitfalls of extrapolations from in vitro to in vivo.
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