A Method for Identifying Mouse Pancreatic Ducts

Tissue Eng Part C Methods. 2018 Aug;24(8):480-485. doi: 10.1089/ten.TEC.2018.0127.

Abstract

Proper identification of pancreatic ducts is a major challenge for researchers performing partial duct ligation (PDL), because pancreatic ducts, which are covered with acinar cells, are translucent and thin. Although damage to pancreatic ducts may activate quiescent ductal stem cells, which may allow further investigation into ductal stem cells for therapeutic use, there is a lack of effective techniques to visualize pancreatic ducts. In this study, we report a new method for identifying pancreatic ducts. First, we aimed to visualize pancreatic ducts using black, waterproof fountain pen ink. We injected the ink into pancreatic ducts through the bile duct. The flow of ink was observed in pancreatic ducts, revealing their precise architecture. Next, to visualize pancreatic ducts in live animals, we injected fluorescein-labeled bile acid, cholyl-lysyl-fluorescein into the mouse tail vein. The fluorescent probe clearly marked not only the bile duct but also pancreatic ducts when observed with a fluorescent microscope. To confirm whether the pancreatic duct labeling was successful, we performed PDL on Neurogenin3 (Ngn3)-GFP transgenic mice. As a result, acinar tissue is lost. PDL tail pancreas becomes translucent almost completely devoid of acinar cells. Furthermore, strong activation of Ngn3 expression was observed in the ligated part of the adult mouse pancreas at 7 days after PDL.

Keywords: Ngn3; differentiation; partial duct ligation; stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cholic Acids / metabolism
  • Fluoresceins / metabolism
  • Fluorescent Dyes / metabolism
  • Ligation
  • Mice, Inbred C57BL
  • Pancreatic Ducts / physiology*
  • Tissue Engineering / methods*

Substances

  • Cholic Acids
  • Fluoresceins
  • Fluorescent Dyes
  • cholyl-lysylfluorescein