Precision cut tissue slices of the liver as morphological tool for investigation of apoptosis.
BACKGROUND
Apoptosis, programmed cell death, is involved in a broad range of pathological processes. Dysregulation of apoptosis plays a key role in the pathogenesis of hepatitis, toxic liver disease and also liver tumor development. For the study of apoptosis in liver diseases, different in vivo models and different in vitro approaches have been developed. They include cell culture models based on hepatocellular carcinoma cell lines or isolated primary hepatocytes.


METHODS
We have established precision cut tissue slices (PCTS) of the liver as a morphological tool for the study of apoptosis. From porcine livers, PCTS were prepared and incubated in a static system with different types and amounts of media. Viability, morphology, spontaneous apoptosis and proliferation were investigated. Apoptosis was induced with actinomycin D and tumor necrosis factor (TNF) alpha.


RESULTS
Morphology and viability was well preserved for at least 24 h. After 48 h, deterioration with single and group cell autolysis was seen. There was a low rate of spontaneous apoptosis and proliferation. Using a combination of TNF alpha and actinomycin D, a significant amount of apoptosis occurred.


CONCLUSIONS
PCTS can be used to directly analyse apoptosis at the tissue level in a qualitative and quantitative manner.
Kasper
Hans-Udo
HU
Institute of Pathology, University of Cologne, Joseph-Stelzmann-Str. 9, D-50931 Koeln, Germany. hans-udo.kasper@uni-koeln.de
Dries
Volker
V
Drebber
Uta
U
Kern
Michael Andre
MA
Dienes
Hans Peter
HP
Schirmacher
Peter
P
D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation; D002470:Cell Survival; D022781:Hepatocytes / Q000648:ultrastructure*; D008099:Liver / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D013552:Swine; D014021:Tissue Preservation