Pancreatic Islets Isolation through Collagenase Perfusion: An Enzymatic Method to Isolate Pancreatic Islet Cells

- Islet cells in the pancreas produce essential hormones involved in endocrine functions. To isolate islets, first, secure a euthanized mouse and surgically expose its abdominal cavity. Brush aside the liver and intestines to locate and clamp the common bile duct. Identify the ampulla of Vater, the duct formed by the union of pancreatic and common bile ducts.

Next, inject a collagenase enzyme solution into the common bile duct. The backflow from the duct successfully inflates the pancreas, facilitating collagenase-induced dissociation of pancreatic tissues. Excise the pancreas into a tube containing more collagenase solution. Mechanically chop the tissue and incubate to aid tissue digestion by the collagenase.

Once obtaining a homogeneous suspension, stop the enzymatic digestion by adding a solution containing proteins that inhibit collagenase activity. Centrifuge to gather a tissue pellet. Resuspend the pellet in an appropriate density gradient medium, layer a suitable buffer over it, and centrifuge to generate gradient layers.

Collect islets from the layer formed between the density gradient medium and the buffer. Finally, pick healthy islets for culturing. In the following proto