Primary human hepatocytes--a valuable tool for investigation of apoptosis and hepatitis B virus infection.
OBJECTIVE
Apoptosis is a key event in the pathophysiology of many liver diseases. Primary human hepatocytes (PHH) provide a useful model to study physiological and pathophysiological processes in the liver. Our aim was to optimize PHH cultures to allow studies on induction of apoptosis and of hepatitis B virus (HBV) infection.


METHODS
PHH were isolated from human liver tissue by two-step collagenase perfusion. PHH and hepatoma cells were treated with different apoptosis-inducing agents in parallel. PHH cultures were infected with wild type HBV and transduced with HBV genomes using adenoviral vectors.


RESULTS
PHH were successfully isolated from 40 different tissue samples with high viability and purity. Perfusion time and seeding density turned out to be critical parameters for optimal cell yield and culture conditions, respectively. Serum addition to the medium reduced viability of PHH. PHH allowed reproducible studies of CD95-dependent and -independent apoptosis. Sensitivity towards CD95-mediated apoptosis was markedly higher than in hepatoma cells. PHH could efficiently be infected with HBV, but infection did neither induce apoptosis nor prevent CD95-induced cell death.


CONCLUSIONS
Our data show that PHH provide an excellent tool for the investigation of apoptosis induced by agents like death receptor-ligands and hepatotropic viruses.
Schulze-Bergkamen
Henning
H
Department of Internal Medicine IV, University of Heidelberg, Heidelberg, Germany.
Untergasser
Andreas
A
Dax
Andreas
A
Vogel
Heiko
H
Büchler
Peter
P
Klar
Ernst
E
Lehnert
Thomas
T
Friess
Helmut
H
Büchler
Markus W
MW
Kirschfink
Michael
M
Stremmel
Wolfgang
W
Krammer
Peter H
PH
Müller
Martina
M
Protzer
Ulrike
U
D017209:Apoptosis / Q000502:physiology*; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000503:physiopathology; D016923:Cell Death; D002469:Cell Separation; D002478:Cells, Cultured; D006509:Hepatitis B / Q000503:physiopathology*; D022781:Hepatocytes / Q000502:physiology* / Q000821:virology*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000503:physiopathology; D019014:fas Receptor / Q000502:physiology
D019014:fas Receptor