Isolation and Culture of Pancreatic Islets

Mice and rats were anesthetized with CO₂ and then killed by decapitation. Their abdomens were opened to expose the pancreas. Subsequently, in situ ductal perfusion was performed. Approximately, 5 and 15 ml collagenase solution (1.2 mg/ml; Roche, Basel, Switzerland) were injected into mouse and rat pancreas, respectively, through the common bile duct. The inflated pancreas was digested by shaking in collagenase solution for 20 min at 37 °C. Then, the digested pancreas was disintegrated by pipetting through a 5-ml pipette tip and rinsed with Hanks balanced salt solution (Invitrogen, Carlsbad, CA, USA) [28 (link), 29 (link)]. The harvested islets were hand-picked and some of them were dispersed into single islet cells. Thereafter, both islets and dispersed islet cells were subjected to cultivation.