Establishment of sandwich cultures of primary human hepatocytes

Scope of the method

Alternative method relates to
  • Human health
Alternative method is situated in
  • Basic Research
Type of alternative 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
Pharmaceutical and Pharmacological Sciences (FARM)