Establishment of sandwich cultures of primary human hepatocytes

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
Primary human hepatocytes


Method keywords
  • Sandwich cultures of hepatocytes
Scientific area keywords
  • Drug-induced cholestasis
Method description

This method describes a well-known optimised human in vitro model of drug-induced cholestasis. Cryopreserved primary human hepatocytes are cultured between two layers of extracellular matrix scaffold, which will delay dedifferentiation and allows to restore cell-extracellular matrix interactions. The sandwich culture method can be applied to both single cell culture dishes and multi-well plates, thus providing an opportune model for high-throughput screening.

Method status
  • Still in development

Pros, cons & Future potential

  • Suitable for long-term exposure;
  • Restored cell polarity;
  • Presence of cell-ECM interactions;
  • Formation of functional bile canalicular network;
  • Maintain functional expression levels of transport proteins and xenobiotic metabolization enzymes;
  • Applicable for quantifying and detecting cholestatic liabilities.
  • Mass transfer barrier;
  • Difficult to culture in 96-well plates;
  • Require daily medium renewal due to accumulating toxic metabolites;
  • Hypoxic environment.

The model is already modified by introducing a renewal of the collagen layer every 3-4 days. As a result, the model shows and extended cultivation regime up to 14 days (Parmentier et al. 2013).

Future & Other applications

The model could be used to assess the overall hepatotoxic potential of drugs, cosmetics, biocides or food additives.

References, associated documents and other information


Gijbels E., Vilas-Boas V., Deferm N. et al. (2019) Mechanisms and in vitro models of drug-induced cholestasis. Archives of Toxicology (submitted) 

Gijbels E., Vanhaecke T., Vinken M. (2019) Establishment of sandwich cultures of primary human hepatocytes. Methods in Molecular Biology - Protocols in Experimental Cholestasis Research (accepted)

Other references you can find in attached document

Associated documents

Contact person

Eva Gijbels


Vrije Universiteit Brussel (VUB)
Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology
Brussels Region