2015 ACVIM Forum Research Abstract Program

Cats that died or were euthanized because of end-stage disorders other than pancreatic disease were included in the study. Initial tests evaluated the best pancreatic infusion technique. Pancreata from nine cats were infused within one hour from death with 60 ml Collagenase Type IV (2 mg/ml, Worthington) through the major duodenal papilla. The perfused pancreas was then digested in a shaking water bath at 37°C. The degree of dissociation of islets from acinar tissue was checked every 5 min by examination with dithizone which specifically stains Zn-complexes in beta cells under a phase-contrast microscope. Collagenase digestion was stopped when nearly 50% of islets were free of acinar tissue. Further purification of islets was achieved by using a filtration method and discontinuous density gradient solutions (1.077 and 1.100 g/cm3; Biochrom). The quality and purity of the final islet product was assessed by DTZ staining. Islet viability was assessed by visual assessment of the membrane integrity of islet cells after double fluorescent staining with fluorescein diacetate and propidium iodide (FDA/PI). In vitro functional activity of purified islets was evaluated by measuring the amount of insulin release (Mercodia Feline Insulin) upon glucose stimulation.