Midline Laparotomy Surgery in Recipient Mice for Orthotopic Liver Transplantation

After shaving and disinfecting the surgical site of an anesthetized recipient mouse, use a midline laparotomy to expose the proper hepatic artery, or PHA. Once the liver is mobilized, ligate the left phrenic and paraesophageal vasculature with a 10-O nylon suture. Next, retract the liver inferiorly and dissect the supra hepatic inferior vena cava, or SHIVC, from the retroperitoneum.

Retract the liver superiorly and dissect the infra hepatic inferior vena cava, or IHIVC, from the retroperitoneum. Then ligate the right adrenal vein and expose the liver hilum. Ligate the PHA with 10-O nylon suture.

Dissect the common bile duct, or CBD, from the portal vein, or PV, and ligate it with a 7-O suture near the bifurcation, creating adequate space for the CBD anastomosis. Once the IHIVC is clamped, temporarily ligate the PV with a 7-O suture. Using a 30 gauge needle, flush 0.5 milliliters of heparin saline into the native liver's portal vein.

Once the SHIVC and IHIVC are clamped, cut the native SHIVC, IHIVC, PV, and any remaining ligamentous attachments to the recipient's native liver. Place the donor liver allograft in the abdominal cavity and retract the hilum of the donor allograft to expose the PV.Once the donor and native PV are flushed, insert the previously made cuff of the donor PV into the lumen of the recipient liver PV.After securing the anastomosis, perform a hand sewn anastomosis using a 10-O nylon suture with the donor and native SHIVC. Finally, complete anastomosis of the superior wall.

Then remove the ligation suture of the PV, followed by the vascular clamps from SHIVC to begin reperfusion. Perform hand sewn anastomosis for IHIVC as demonstrated for SHIVC. After completing the reconstruction, remove the micro clamp or suture from the IHIVC to reperfuse.

Once a ductotomy is created, insert the donor CBD stent into the recipient CBD lumen and secure the anastomosis with a 10-O suture.