The establishment of reperfusion system in isolated mouse liver

Objective To establish the reperfusion system in isolated mouse liver and test its effectiveness. Methods The isolated livers of 6 male C57 mice were reperfused in vitro with cold preservation time less than 45 minutes, and the blood obtained from allogenic mice was made up into perfusate before reperfusion. The power of reperfusion system came from peristaltic pump, and a dripping bucket was added for stress-buffering and venting. Two animal membrane oxygenation were in series with the reperfusion system to meet oxygen requirements. The total liquid of reperfusion system was about 30 mL, and the rate of reperfusion was set as 2.5 mL·min-1·g-1. Related parameters, such as liver perfusion pressure were recorded by biological functional experiment system, and indexes like ALT, AST of perfusate, histopathology of livers and cell apoptosis were also monitored. Results During reperfusion in vitro, the portal vein pressure was lower than 8 mmHg, the system pressure fluctuation was lower than 1 mmHg, and the pH of the perfusion fluid was 7.40±0.05. At the end of reperfusion, the concentration of ALT, AST and lactate dehydrogenase of perfusion fluid were (62±14), (160±23) and (682±71) U/L, respectively, the concentration of lactic acid was (3.2±0.1) mmol/L. The pathology results of liver tissue showed that the structure of hepatic lobule was intact without obvious necrosis or apoptosis hepatocyte, and the pathological score was (1.1±0.7) scores. The apoptosis rate of hepatocyte was (0.57±0.22)%. Conclusions For the first time, the isolated reperfused mouse liver system was constructed. It is simple, stable, and could maintain normal physiological function of the liver for 6 hours. Key words: Mouse; Isolated perfused system; Liver