ERM 324 (UP, CHU et INSERM). Ischemia-reperfusion during organ transplantation and improvement of organs conservation conditions. 18 researchers.
Organs to be transplanted are usually conserved at 4°C in solutions highly concentrated in potassium which can lead to damageable lesions in the long term. Shortages of donors however means that surgeon have to use organs even though their conservation conditions are considered unsatisfactory.
Research topics:
to improve general organ conservation conditions and to enable the use of organs which are not in the best condition for transplants. For this purpose two approaches are considered:
The in vivo / ex vivo approach. During kidney auto-transplants new conservation solutions are tested: they contain polyethylene glycol and a saline solution mimicking extra-cellular conditions as base components, and are complemented by candidate molecules (studied in vitro, see below) with a potential to act on biochemical mechanisms. High-resolution magic angle spinning (HRMAS) NMR spectrometry wide usage should lead to an application for humans.
The in vitro approach. The triggering of cell death and the apoptotic stages is monitored in hypoxic and re-oxygenation conditions respectively by titrating caspases, measuring phosphatidylserine surface exposure by immunostaining, by TUNEL assay and propidium iodide staining. The following biochemical parameters are also studied: phosphatidylinositol 3-kinases, lipid oxidation (plasmalogens and the production of aldehyde, isoprostane, and PAF-like), modifications of the cytoskeleton, gene expression (HIF, cytokines, adhesion molecules, PBR), mitochondrial respiration and ATP production in isolated tubular cells and in cultured cells.
In collaboration with a local company Bio-alternatives, development of cDNA chips as a diagnostic and prognostic tool (differential display studies in progress)
Techniques and equipment of interest:
Kidney auto-transplantation in pigs with different methods of organ conservation
Kidney ischemia-reperfusion in rats and pig by clamping of arteries and blood vessels.
Spectroscopy by proton nuclear magnetic resonance, C13 and P31 on biological fluids & biopsies.
Overexpression of genes in cell cultures.
Multi-technologies plate analyser: visible-UV absorption, fluorescence, luminescence
Capillary electrophoresis
Liquid & gas chromatography coupled with mass spectroscopy
