Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

We focus our research on liver transplantation, in particular on cold ischemia and warm-reperfusion injury. We use mouse liver transplantation model as a research tool for this purpose. Because the technique is difficult, so it can take a long time to learn and to achieve consistent results.

In the future, we'd like to conduct research on fatty liver transplantation. This is still a problem to overcome on clinical liver transplantation. To begin, prepare 16 gauge and 20 gauge intravenous catheters for the portal vein, or PV, and the infrahepatic inferior vena cava, or IHIVC respectively.

Using a scalpel, cut the catheter to create the cuff. Then with the back of the scalpel, create a shallow groove on the cuff's surface for thread fixation. Place small ice cubes in a rectangular-shaped plastic container, and position a small metal cup on top of the container.

Fill the cup with organ preservation fluid and place the syngeneic liver graft in it. Under eight to 10X magnification, gently roll the liver graft using a cotton swab, ensuring that the porta hepatis faces upwards. Thread the PV through the cuff using a thread attached to the splenic vein.

With the cuff handle at the 12 o'clock position, secure the handle and cuff with a bulldog clamp, positioning it two to three millimeters from the PV edge. Further, secure the bulldog clamp using large vascular forceps and a fixation mold to establish the operative field. Next, increase the magnification to 15 to 20X and carefully inspect the PV lumen.

After grasping the PV lumen, externalize the thin wall of PV through the inner lumen of the cuff. Secure the fully externalized PV lumen with a double ligation using 8-O silk thread along the cuff's groove, and then cut the string. Similarly, attach the cuff to the IHIVC lumen and fix it using 8-O silk thread.

Then to create a slipknot, pass the thread around the base of the IHIVC to temporarily ligate it. After extracting the native liver from the anesthetized recipient mouse, gently grasp the recipient's liver portal vein, or PV, edge using pean forceps and secure it with a vice. Pass an 8-O silk thread around the PV and place a vascular clamp on it.

Create a one fourth circumferential section approximately 0.5 millimeters from the PV edge. While passing saline through the hole, use a specialized instrument to enlarge the hole. Grip the cuff handle using straight forceps and insert it into the lumen.

Then secure the cuff with an 8-O silk thread. Remove the vascular clamp from PV to allow reperfusion of the liver. Finally, carefully release the pean forceps to complete the PV reconstruction.

After positioning the liver lobe, move the pean forceps attached to the recipient IHIVC dorsal to the graft's right lower lobe. Then secure the IHIVC with a vice. Rub away any remaining liver tissue around the recipient's IHIVC with a cotton swab.

Pass an 8-O silk thread around the recipient's IHIVC. Create a one fourth circumferential section approximately 0.5 millimeters from the recipient's IHIVC edge. After flushing the lumen with saline, insert the cuff into the IHIVC lumen.

Carefully secure the cuff with an 8-O silk thread. Finally, remove the vascular clamp and pean forceps. The preservation of the graft at cold temperatures for one hour resulted in serum alanine aminotransferase levels averaging approximately 2, 000 units per liter at six hours post reperfusion.

A survival rate of 100%was observed at seven days following transplantation.