Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Abstract

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

TRPV4 channel opening mediates pressure-induced pancreatitis initiated by Piezo1 activation

“…Histological scoring was evaluated from the entire section. Partial PDL was performed as previously described (89,90). Using a stereo microscope, the tail region of the pancreas was visualized, and the main pancreatic duct was ligat-the appropriate level of PLA2 activity.…”

Pancreatic duct ligation reduces premalignant pancreatic lesions in a Kras model of pancreatic adenocarcinoma in mice

“…At 12 weeks of age, the peritoneal cavity was accessed through a midline laparotomy, under anesthetic induction with isoflurane at 4% and at 2.5% maintenance in 100% oxygen. The stomach, pancreas and spleen were mobilized as described previously 19 , 24 in all the animals subjected to surgery (sham and PDL groups). In PDL group mice, dissection and ligation of the splenic lobe of the pancreas were performed on the right side of the superior mesenteric vein, using 8/0 polyglycolic acid suture (Fig.…”

Semi-automated digital measurement as the method of choice for beta cell mass analysis

“…All animal experiments were performed according to the guidelines of our institutional "Ethical Committee for Animal Experiments" (ECD 12-595-5) and national guidelines and regulations. Pancreatic partial duct ligation or Sham operation was performed as described [ 1 , 11 ] on 8 weeks old, male Balb/c mice.…”