Establishment of sandwich cultures of primary human hepatocytes
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- Sandwich cultures of hepatocytes
- Drug-induced cholestasis
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.
- 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).
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
Contact personEva Gijbels
OrganisationsVrije Universiteit Brussel
Pharmaceutical and Pharmacological Sciences (FARM)
In Vitro Toxicology and Dermato-cosmetology