Assessment of the cholestatic potential of drugs using primary human hepatocyte spheroids
Scope of the method
- Human health
- Translational - Applied Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- primary human hepatocytes
- ATP quantification
- cholestatic index
- in vitro toxicology
- cholestatic liability
This method describes a very reliable and robust in vitro model for the screening of the cholestatic liability of drugs and other chemical entities. The 3D spheroids generated from primary human hepatocytes can be cultivated up to 28 days, allowing long-term exposures which can depict otherwise undetectable toxicity. After spheroid generation, these are exposed to the test drug in the presence or in the absence of a 30x concentrated mixture of the 5 most relevant bile acids (BA) up to 28 days, with medium renewal every 2 days. At the pre-established time-points, cell viability is checked using an ATP quantification kit and the cholestatic potential of the studied drug is determined by means of the Cholestatic Index (CIx). This is calculated as the ratio between the ATP content in spheroids treated with drug plus BA and the spheroids treated only with the drug. A CIx equal or below 0.8 suggests increased cholestatic liability for the tested drug.
- Biosafety cabinet ;
- Centrifuge with rotor for plates ;
- Microscope ;
- Incubator at 37ºC with controlled atmosphere ;
- Plate reader for bioluminescence.
- Still in development
Pros, cons & Future potential
- Increased similarity to the in vivo situation ;
- Allows studying long-term effects.
- Expensive methodology.
The method has not yet been tested with chemicals other than drugs, so the IVTD group is further studying this possibility.
References, associated documents and other information
Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease. Bell CC et al. Sci Rep. 2016 May 4;6:25187. doi: 10.1038/srep25187
Three-Dimensional Spheroid Primary Human Hepatocytes in Monoculture and Coculture with Nonparenchymal Cells. Baze AC et al. Tissue Eng Part C Methods. 2018 Sep;24(9):534-545. doi: 10.1089/ten.TEC.2018.0134
Contact personVânia Vilas-Boas
OrganisationsVrije Universiteit Brussel (VUB)
Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology