Heparin sodium 25000

All surgical interventions were performed under inhalation anesthesia (2 vol. % isoflurane mixed with 0.3 L/min oxygen using an isoflurane vaporizer, Sigma Delta, UK) after analgesic pretreatment with buprenorphine (Temgesic®; 0.05 mg/kg s.c.) applied 0.5 h prior to the skin incision. Briefly, after the induction of anesthesia, systemic heparinization (300 IU/kg body weight Heparin-sodium-25000-ratiopharm®, Ratiopharm, Germany) was performed. After completion of the respective operative procedure, animals were placed on a heating pad during the recovery period. For postoperative analgesic treatment, buprenorphine was applied subcutaneously twice daily for 3 postoperative days at a dose of 0.05 mg/kg body weight.

After placing the anesthetized animals in the supine position, the abdomen was opened via a wide transversal incision. The liver was mobilized by dissecting the triangular ligament and the portal vein was exposed. Two ligatures were placed 1 cm below the liver hilum on the portal vein. The portal vein was cannulated with a 26 gauge catheter (Terumo Europe®, Belgium) that was fixed with two suture lines. Total heparinization was achieved by slowly injecting 300 IU heparin/kg body weight; (Heparin-sodium-25000-ratiopharm®, Ratiopharm, 25000 I.E.) via the portal vein over a period of 10 minutes using a non-peristaltic pump (0.3 ml/min; Perfusor VI, B. Braun, step 7). The diaphragm was opened to place an incision on the suprahepatic vena cava, which allowed the drainage of the perfusate.
After total perfusion of the liver, the organ was freed from the remaining ligaments and removed from the abdominal cavity. This was directly followed by taking a native liver tissue sample, preferably from the inferior caudate liver lobe.