Pancreatic islets (cadavers) (University of Alberta, Canada)

To isolate the islets, the ducts were perfused, in a controlled fashion, with a cold enzyme (Liberase human islet, Roche). The islets were then separated, by gentle mechanical dissociation, and purified with the use of continuous gradients of Ficoll–diatrizoic acid (Seromed-Biochrom) in an apheresis system. To minimize the risk of islet injury due to cold ischemia, freshly prepared islets were transplanted immediately after harvesting, thus eliminating the need for islet culture. The islets were incubated for 24 hours at 37°C in CMRL 1066 medium with fetal-calf serum (10%) and HEPES buffer (25 mmol).