Improving the preservation of the non-heart-beating-donor pancreas

• Main Author: Reddy, Mettu
• Published: University of Sunderland 2010
• Subjects: 617; Health Sciences
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.542250

Successful pancreatic islet transplantation depends on the number of islets transplanted, islet viability and extent of early islet loss due to ischaemia reperfusion injury. Novel pancreas preservation techniques which can improve these variables can increase the utilisation of non-heart-beating-donor pancreases for islet transplantation. A non-heart-beating-donor rat pancreas preservation model was developed. Pancreases preserved by either static cold storage, hypothermic machine perfusion or portal venous oxygen persufflation underwent islet isolation and purification. The yield, viability and in vitro function of isolated islets were compared. Portal venous oxygen persufflation improved the islet yield, viability and morphology as compared to static cold storage. The percentage of pancreases with functional islets (stimulation index greater than 1.0) was also higher after oxygen persufflation as compared to static cold storage. Severity of reperfusion injury in pancreases preserved by static cold storage or portal venous oxygen persufflation was compared after in vitro warm reperfusion. Amylase, lipase and glycerol levels in the portal effluent were measured. Lipid peroxidation and apoptosis in reperfused pancreas were measured using thio-barbituric acid reactive substances assay and caspase 3 assay respectively. Expression of genes relevant to ischaemia reperfusion was compared using RNA microarrays. Severity of ischaemia reperfusion injury was similar in both groups. Microarray analysis revealed increased expression of genes related to apoptosis in the portal venous oxygen persufflation group. This group also showed up-regulation of pro-survival cellular pathways and over-expression of genes related to cellular repair as compared to static cold storage. This project has for the first time evaluated oxygen persufflation as a method of pancreas preservation and investigated changes in global gene expression after this form of preservation. Overall, the project suggests that portal venous oxygen persufflation improves the recovery of non-heart-beating-donor rat pancreas. Further investigation to examine its role in the preservation of large animal pancreases is needed.