Pancreatic intra-ductal lentivirus infusion in mice

The procedure for retrograde infusion of lentivirus into pancreatic duct has been described by Xiao et al³⁶. Briefly, the male C57BL/6J mice (8 weeks old, about 23–25 g) were anesthetized with isoflurane. A midline incision was made to reveal the abdominal cavity. Q-tips were used to gently pull out the stomach. Rotate and stretch the duodenum to expose the biliary-pancreatic duct and its junction with the duodenum. A microclamp was placed on the bile duct above the branching of the pancreatic duct to prevent perfusion of the liver. Then a 30-gauge needle was inserted through the anti-mesenteric aspect of the duodenum to cannulate the common bile duct and the tip of the catheter should be positioned at the origin of the pancreatic duct branch in the biliary-pancreatic duct. A small microclamp was applied to the distal common bile duct to prevent back flow of the infusate into the duodenal lumen and to hold the cannula in place. 1 × 10⁹ indicated lentivirus dissolved in 0.15 ml normal saline (NS) or normal saline (NS) was infused at 6 µl per min for 25 min using a R462 perfusion pump (RWD, China, shengzhen). After infusion, close the hole in the duodenum created by the catheter with a 7-0 suture. Upon completion of the infusion, the clamps were released. The exterior abdominal wound was closed using 7 mm wound clips. Mice recovered on a heating pad (37 °C) until it fully recovers. They were given free access to food and water after the surgery. At 72 h after injection, islets were lysed to extract total RNA or protein to measure the overexpression efficacy. At 1 week after injection, mice were fed with high fat diet (HFD, {"type":"entrez-nucleotide","attrs":{"text":"D12494","term_id":"217719","term_text":"D12494"}}D12494, 60% energy from fat) for 16 weeks.