Murine Model of Islet Transplantation: A Surgical Method to Infuse Pancreatic Islets in Portal Vein and Prevention of Postoperative Bleeding
Transkript
In a mouse model of insulin-dependent diabetes, pancreatic islets are injected via the hepatic portal vein for therapeutic purposes. The beta cells within the islet cluster produce insulin, a hormone that regulates blood glucose levels.
Intraportal injection is a simple and effective technique, but postoperative bleeding often leads to the death of the model animal. To prevent this complication, prep an anesthetized mouse in the supine position. Shave the abdominal area and apply a disinfectant solution. Make an incision to expose the peritoneal cavity. Exteriorize the intestines from the peritoneal cavity on a moist gauze to expose the underneath portal vein.
Take an insulin syringe pre-loaded with islets. Insert the needle directly parallel into the portal vein, with its bevel facing down. Draw some blood into a syringe to mix the islets, and then inject slowly. Pull and push the plunger multiple times to infuse the entire islet suspension. Now, place a gelatin sponge to cover the injection site.
Gelatin, a hemostatic matrix, absorbs blood and swells to form a stable clot over the puncture site, thereby arresting the bleeding. Next, return the intestine into the peritoneal cavity and suture the incision. Return the mouse to its home cage and regularly monitor for signs of recovery.
