Isolation and Culture of Pig Hepatocyte in Large Scale for the Application of Bioartificial Liver System / 대한간학회지

BACKGROUND/AIMS: Acute hepatic failure is a serious problem. Its mortality reaches up to 80%. Only liver transplantation has been accepted as a definite treatment for patients with hepatic failure but shortage of donor organs is the main obstacle of this approach. A possible solution to this problem is a bioartificial liver system, perfusion of patients blood to isolated hepatocyte. In this study, we performed the isolation and culture of pig hepatocyte in large scale for the application of bioartificial liver system. METHODS: Hepatocyte isolation was performed by two-step collagenase method via portal vein perfusion in 10kg female pigs. After that, we compared the functional differences of the spheroid culture to the monolayer culture of hepatocyte. The viability and the function of hepatocyte were assessed using trypan-blue exclusion test and the measurement of the rate of ureagenesis and ammonia removal. RESULTS: The average viability and yield of hepatocyte were 86.8 +/- 8.0 % and 7.8 +/- 5.4 X 10(9), respectively. The spheroid culture was superior to the monolayer culture in functional aspect of hepatocyte, and their differences, especially for ammonia removal, were more apparent in parallel with culture time. CONCLUSIONS: For hepatocyte isolation, we obtained sufficient viability and yield of hepatocyte for clinical usage of bioartificial liver system. The function of hepatocyte seems to be better in the spheroid culture than in the monolayer culture. Further studies are needed for application of bioartificial liver system in clinical setting.

Bioartificial liver system for treatment of acute liver failure in canines and the effect on serum endotoxin / 中国普通外科杂志

Objective To evaluate the effect of a bioartificial liver (BAL) system for treatment of acute liver failure (ALF) in canines and its relationship to serum endotoxin. Methods ALF was induced by end-side portocaval shunt combined with common bile duct ligation and transection. Ten ALF canines were distributed to BAL group (n=5), or to a control group (n=5). Each BAL circulation lasted 5h. Serum endotoxin, alanine aminotransferase (ALT) and total bilirubin (TB) before establishment of ALF model, pre-circulation and post-circulation were determined. Results Serum endotoxin level was 0.284 EU/ml, 0.526 EU/ml and 0.416 EU/ml before establishment of ALF model, and pre-circulation and post-circulation in BAL group, respectively. The serum endotoxin level in BAL group increased pre-circulation (P

Rat-Hepatocyte Culture and Differentiation in Hormone-Supplemented Media

BACKGROUND: Severe liver disease is very often life threatening and dramatically diminishes the quality of life. An effective artificial liver support system should be capable of sustaining patients with severe liver disease, preparing patients for liver transplantation, and improving the survival of and the quality of life for patients for whom transplantation is not a therapeutic options. METHODS: As an elemental experiment for establishing a bioartificial liver system, rat hepatocytes were isolated in several hormone-supplemented culture media: 1) control group, 2) insulin supplemented media, 3) EGF supplemented media, 4) insulin EGF supplemented media. Using primarily the cultured and subcultured hepatocytes, we evaluated the hepatic cellular function in each media. RESULTS: The cellular viability was maintained as long as 10 days (average, 8.4 days). The hepatocytes in the group 4 media showed good results in ammonia degradation, ureogenesis, and albumin synthesis compared with those in the other groups. In the subculture, the hepatocyte functions had significantly declined. CONCLUSION: Insulin EGF supplemented culture media were superior to the others.