Midline Laparotomy Surgery in Donor Mice for Orthotopic Liver Transplantation

To begin, place the anesthetized donor mouse under the high-powered microscope. Using a pair of scissors, perform a midline laparotomy from the xiphoid process to the pubic synthesis. Then make a transverse incision below the ribs to create a cross like pattern.

After retracting the xiphoid process, eviscerate the intestines, and place them on the wet gauze sponge on the left side of the abdominal cavity. To mobilize the liver, separate it from all its ligamentous attachments. Once the proper hepatic artery or PHA is exposed, using curved forceps, skeletonize it.

After ligating the PHA, dissect the entire length of the common bile duct, or CBD. Insert the bile duct stent in the CBD and secure it with a 10-O nylon suture, then ligate the distal aspect of CBD. Using a wet gauze sponge, retract the right hepatic lobe toward the xiphoid and expose the inferior vena cavo, or IVC.

Mobilize the infra hepatic inferior vena cava, or IHIVC, away from the retroperitoneum and cauterize the right adrenal vein. Once the right renal artery and vein are dissected and ligated, cut both and the remaining ligamentous attachments. After removing the right kidney, inject 0.5 milliliters of cold histidine trytophan ketoglutarate solution with 100 units of heparin through the portal vein, or PV.After one minute of injection, cut the portal vein superior to the splenic and superior mesenteric veins.

To perfuse the liver, slowly inject a cold histidine trytophan ketoglutarate preservation solution into the IHIVC until the fluid coming from the PV is clear. Place a micro clamp or fine suture on the IHIVC just superior to the right renal vein, and cut just inferiorly to the clamp. Once the CBD is cut distally to the stent, identify and ligate the cystic duct with a 10-O nylon suture.

With forceps, grasp the dome of the gallbladder, and using spring scissors, cut the cystic duct above the suture to complete the cholecystectomy. Retract the liver inferiorly to expose the supra hepatic inferior vena cava, or SHIVC, and cut it, preserving an adequate length for the anastomosis in the recipient mice. After removing the additional ligamentous attachments.

Place the liver in cold saline.