Bile canalicular dynamics in hepatocyte sandwich cultures
Journal article, 2015

Many substances are hepatotoxic due to their ability to cause intrahepatic cholestasis. Therefore, there is a high demand for in vitro systems for the identification of cholestatic properties of new compounds. Primary hepatocytes cultivated in collagen sandwich cultures are known to establish bile canaliculi which enclose secreted biliary components. Cholestatic compounds are mainly known to inhibit bile excretion dynamics, but may also alter canalicular volume, or hepatocellular morphology. So far, techniques to assess time-resolved morphological changes of bile canaliculi in sandwich cultures are not available. In this study, we developed an automated system that quantifies dynamics of bile canaliculi recorded in conventional time-lapse image sequences. We validated the hepatocyte sandwich culture system as an appropriate model to study bile canaliculi in vitro by showing structural similarity measured as bile canaliculi length per hepatocyte to that observed in vivo. Moreover, bile canalicular excretion kinetics of CMFDA (5-chloromethylfluorescein diacetate) in sandwich cultures resembled closely the kinetics observed in vivo. The developed quantification technique enabled the quantification of dynamic changes in individual bile canaliculi. With this technique, we were able to clearly distinguish between sandwich cultures supplemented with dexamethasone and insulin from control cultures. In conclusion, the automated quantification system offers the possibility to systematically study the causal relationship between disturbed bile canalicular dynamics and cholestasis.

Cholestasis

Bile canaliculi

3D system

Collagen sandwich

Primary hepatocytes

Image quantification

Author

Raymond Reif

Johan Karlsson

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Georgia Günther

Lynette Beattie

David Wrangborg

Seddik Hammad

Brigitte Begher-Tibbe

Amruta Vartak

Simone Melega

Paul M. Kaye

Jan G. Hengstler

Mats Jirstrand

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Archives of Toxicology

0340-5761 (ISSN) 1432-0738 (eISSN)

Vol. 89 10 1861-1870

Areas of Advance

Information and Communication Technology

Life Science Engineering

Subject Categories

Computational Mathematics

Bioinformatics and Systems Biology

Roots

Basic sciences

DOI

10.1007/s00204-015-1575-9

PubMed

26280096

More information

Created

10/8/2017