All mouse experiments were approved by the Animal Research and Care Committee at the Children’s Hospital of Pittsburgh and the University of Pittsburgh IACUC. BAC transgenic glucagon promoter Cre reporter (GCG-Cre) mice were purchased from MMRRC (Chapel Hill, NC, USA) [30 (link)]. The BAC transgenic elastase promoter CreERT reporter (Ela-CreERT) mouse has been described before [31 (link), 32 (link)]. C57/6, Rosa26CAGTomato (Tomato) and BAC transgenic mouse insulin promoter green fluorescent protein reporter (MIP-GFP) mice [20 (link)] were all purchased from Jackson ImmunoResearch (Bar Harbor, Maine, USA). Tamoxifen induction of Tomato expression in acinar cells in Ela-CreERT; Tomato mice have been described before [32 (link)]. All experiments used 8-week-old male mice. Fasting blood glucose monitoring and intraperitoneal glucose tolerance test (IPGTT) were performed as described previously [20 (link), 33 (link), 34 (link)].
Pancreatic intraductal virus infusion was performed as described previously [32 (link), 35 (link)], but here we delivered 50 μl adeno-associated virus serotype6 vectors (AAV6) (titration of 109) via catheter at a rate of 10 μl/min to optimise infection of duct cells in the current study. Islet transplantation was performed on a heated plate. The left kidney of the mouse was exposed through a lumbar incision. The kidney capsule was then incised with a Hamilton syringe (Fisher Scientific, Pittsburgh, PA, USA) and a pocket was made with polyethylene tubing, after which islets were placed under the kidney capsule. The incision in the kidney capsule was cauterised and the peritoneum and skin were closed with suture.
Pancreatic intraductal virus infusion was performed as described previously [32 (link), 35 (link)], but here we delivered 50 μl adeno-associated virus serotype6 vectors (AAV6) (titration of 109) via catheter at a rate of 10 μl/min to optimise infection of duct cells in the current study. Islet transplantation was performed on a heated plate. The left kidney of the mouse was exposed through a lumbar incision. The kidney capsule was then incised with a Hamilton syringe (Fisher Scientific, Pittsburgh, PA, USA) and a pocket was made with polyethylene tubing, after which islets were placed under the kidney capsule. The incision in the kidney capsule was cauterised and the peritoneum and skin were closed with suture.
