Assessment of the cholestatic potential of drugs using primary human hepatocyte spheroids

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.