Human liver slices as an in vitro model to study toxicity-induced hepatic stellate cell activation in a multicellular milieu.
BACKGROUND
Hepatic stellate cell (HSC) activation is a key event in wound healing as well as in fibrosis development in the liver. Previously we developed a technique to induce HSC activation in slices from rat liver. Although this model provides a physiologic, multicellular milieu that is not present in current in vitro models it might still be of limited predictive value for the human situation due to species-differences. Therefore, we now aimed to evaluate the applicability of human liver slices for the study of HSC activation.


METHODS
Liver slices (8 mm diameter, 250 microm thickness) were generated from human liver tissue and incubated for 3 or 16 h with 0-15 microl of carbon tetrachloride (CCl4) after which ATP-content and expression levels of HSC (activation) markers was determined.


RESULTS
Human liver slices remained viable during incubation as shown by constant ATP levels. Incubation with CCl(4) caused a dose-dependent decrease in viability and an increase in mRNA expression of the early HSC activation markers HSP47 and alphaB-crystallin, but not the late markers for HSC activation, alphaSMA and pro-collagen 1a1. Synaptophysin mRNA expression remained constant during incubation with or without CCl4, indicating a constant number of HSC in the liver slices.


CONCLUSIONS
We developed a technique to induce early toxicity-induced HSC activation in human liver slices. This in vitro model provides a multicellular, physiologic milieu to study mechanisms underlying toxicity-induced HSC activation in human liver tissue.
van de Bovenkamp
M
M
Department of Pharmacokinetics and Drug Delivery, University Center for Pharmacy, Groningen, The Netherlands. Electronic address: M.van.de.Bovenkamp@rug.nl.
Groothuis
G M M
GMM
Department of Pharmacokinetics and Drug Delivery, University Center for Pharmacy, Groningen, The Netherlands.
Meijer
D K F
DKF
Department of Pharmacokinetics and Drug Delivery, University Center for Pharmacy, Groningen, The Netherlands.
Slooff
M J H
MJH
Department of Surgery, Division of Hepatobiliary Surgery and Liver Transplantation, University Hospital Groningen, The Netherlands.
Olinga
P
P
Department of Pharmacokinetics and Drug Delivery, University Center for Pharmacy, Groningen, The Netherlands.
D002251:Carbon Tetrachloride / Q000633:toxicity; D002470:Cell Survival / Q000187:drug effects; D050958:HSP47 Heat-Shock Proteins / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008954:Models, Biological; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D038203:alpha-Crystallin B Chain / Q000235:genetics
D050958:HSP47 Heat-Shock Proteins; D012333:RNA, Messenger; C494660:SERPINH1 protein, human; D038203:alpha-Crystallin B Chain; D002251:Carbon Tetrachloride