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

Korean J Hepatol > Volume 8(3); 2002 > Article

The Korean Journal of Hepatology 2002;8(3): 249-255.

원저 : 인공간 시스템 적용을 위한 돼지 간세포의 대량 분리 및 배양 ( Original Articles : Isolation and Culture of Pig Hepatocyte in Large Scale for the Application of Bioartificial Liver System )

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

Yu Jeong Chung2, Hyuk-Joon Lee1, Young Taeg Koh3, Sang Beom Kim1 , Seong Hoon Kim1, Seokho Choi1, Nam-Joon Yi11, Seong-Hwan Chang1, Eun Lan Yang2, Kyung-Suk Suh1, Yoon Shin Lee4 , and Kuhn Uk Lee1,2

Department of Surgery, Seoul National University College of Medicine1 Clinical Research Institute, Seoul National University Hospital2 Department of Surgery, National Police Hospital3 Department of Biomedical Engineering4, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT

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±5.4×10⁹, 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. (Korean J Hepatol 2002;8:249-255)

KeyWords: Bioartificial liver system, Hepatocyte, Monolayer culture, Spheroid culture, Pig