In Vivo Murine Liver Perfusion: A Method to Infuse the Rodent Liver with Solution via the Portal Vein
Transcrição
To place the portal vein catheter, first, use the back of a forceps to move the intestines to the right side of the mouse abdominal cavity, exposing the portal vein, and use curved forceps to slide a 20-centimeter piece of polyester sewing thread under the portal vein between the liver and the superior pancreaticoduodenal vein.
Use the forceps to carefully pull the thread to the other side of the animal so that it is centered under the portal vein and tie a loose overhead knot around the portal vein. Place the curved forceps under the vein and gently pull all tissue material toward the tail to straighten out the vein.
Next, place the bevel of a catheter needle facing up and parallel with the lower part of the portal vein near the forceps and gently puncture the vein with the needle. When the bevel is in the lumen of the vein, retract the spring-loaded needle and continue to push the polymer catheter through the vein until the bevel is near the venous branched area.
After securing the knot around the catheter, immediately set the peristaltic pump at 4 milliliters per minute and connect the male end of the tubing to the female end of the catheter, at which point you should start to see the liver blanch. Cut the aorta abdominalis for drainage. Use masking tape to immobilize the tubing onto the underpad.
Use the scissors to cut the abdominal skin vertically to allow drainage of blood and perfusion liquid. Then, squeeze the effluent blood vessel a few times with straight forceps to inflate the liver, ensuring that all of the blood has drained out, and perfuse the liver with PBS until the liver tissue blanches.
