Human liver slices as an in vitro model to study toxicity-induced hepatic stellate cell activation in a multicellular milieu

Chem Biol Interact. 2006 Jul 25;162(1):62-69. doi: 10.1016/j.cbi.2006.05.006. Epub 2006 May 17.

Abstract

Introduction: 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.

Method: 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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Tetrachloride / toxicity
  • Cell Survival / drug effects
  • HSP47 Heat-Shock Proteins / genetics
  • Hepatocytes / cytology*
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Hepatocytes / pathology
  • Humans
  • In Vitro Techniques
  • Liver / cytology
  • Liver / drug effects*
  • Liver / metabolism
  • Liver / pathology
  • Models, Biological*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • alpha-Crystallin B Chain / genetics

Substances

  • HSP47 Heat-Shock Proteins
  • RNA, Messenger
  • SERPINH1 protein, human
  • alpha-Crystallin B Chain
  • Carbon Tetrachloride