Improvement of hepatocyte recovery from rat liver subjected to 1-hour warm ischemic injury by using citrate phosphate dextrose added to euro-collins perfusion solution

Abstract

Warm ischemia (WI)-related injury interferes with recovery of primary hepatocyte after collagenase digestion of surgically resected or non-heart-beating donor livers as human cell sources. We speculated that digestion is impaired due to reduced microcirculation, caused by microembolism after WI. We sought to improve hepatocyte recovery after WI using a rat model. Anesthetized 9-week-old male Sprague-Dawley rats underwent a midline abdominal incision to insert a 22-gauge cannula into the portal vein. WI was initiated by ligating both the cannula and the hepatic artery. We compared Euro-Collins (EC) perfusion solution with 2 anticoagulants-heparin or citrate phosphate dextrose (CPD)-versus ethylene glycol tetraacetic acid (EGTA) combined with Ca(2+)- and Mg(2+)-free Hank's solution (CM-free Hank's solution). Use of CM-free Hank's solution yielded only 0.75 ± 0.15 × 10(8) and 0.82 ± 0.20 × 10(8) cells at 30 and 60 minutes WI respectively. However, CPD, but not heparin, added to the EC solution produced the best cell recovery (CPD: 2.15 ± 0.38 × 10(8); heparin: 1.63 ± 0.31 × 10(8)). During macroscopic observation, CPD added to EC solution also demonstrated best blood flushing. CPD added to EC solution achieved greater hepatocyte recovery than CM-free Hank's solution by restoring microcirculation during flushing of blood from liver tissue subjected to WI.