A System for ex vivo Culturing of Embryonic Pancreas

The overall goal of this procedure is to dissect and culture mouse embryonic pancreatic buds, ex vivo, allowing direct visualization of the developing pancreas. This is accomplished by first dissecting embryonic day 11.5 or 12.5 mouse embryos and removing the upper body and tail region to isolate the mid body of the embryo. The second step is to expose the duodenum region and the stomach for location, and then dissection of the dorsal pancreatic bud.

Next, the bud is transferred into the micro well of a MacTech dish and cultured for up to one week. Finally, the explan can be analyzed for live cell imaging, or the explan can be fixed, whole mount stained and analyzed by immunofluorescence imaging. Ultimately pancreatic cell differentiation, cytoskeleton organization and cell polarity and branching morphogenesis can be evaluated.