title: [Isolation and culture of pig hepatocyte in large scale for the application of bioartificial liver system].
abstract: OBJECTIVE
  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 10 kg 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.
authors.affiliation: Department of Surgery, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Yu Jeong
authors.initials: YJ
authors.lastname: Chung
authors.forename: Hyuk Joon
authors.initials: HJ
authors.lastname: Lee
authors.forename: Young Taeg
authors.initials: YT
authors.lastname: Koh
authors.forename: Sang Beom
authors.initials: SB
authors.lastname: Kim
authors.forename: Seong Hoon
authors.initials: SH
authors.lastname: Kim
authors.forename: Seok Ho
authors.initials: SH
authors.lastname: Choi
authors.forename: Nam Joon
authors.initials: NJ
authors.lastname: Yi
authors.forename: Seong Hwan
authors.initials: SH
authors.lastname: Chang
authors.forename: Eun Lan
authors.initials: EL
authors.lastname: Yang
authors.forename: Kyung Suk
authors.initials: KS
authors.lastname: Suh
authors.forename: Yoon Shin
authors.initials: YS
authors.lastname: Lee
authors.forename: Kuhn Uk
authors.initials: KU
authors.lastname: Lee
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D005260:Female; D022781:Hepatocytes / Q000166:cytology*; D019164:Liver, Artificial; D013552:Swine