Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease

Sci Rep. 2016 May 4:6:25187. doi: 10.1038/srep25187.


Liver biology and function, drug-induced liver injury (DILI) and liver diseases are difficult to study using current in vitro models such as primary human hepatocyte (PHH) monolayer cultures, as their rapid de-differentiation restricts their usefulness substantially. Thus, we have developed and extensively characterized an easily scalable 3D PHH spheroid system in chemically-defined, serum-free conditions. Using whole proteome analyses, we found that PHH spheroids cultured this way were similar to the liver in vivo and even retained their inter-individual variability. Furthermore, PHH spheroids remained phenotypically stable and retained morphology, viability, and hepatocyte-specific functions for culture periods of at least 5 weeks. We show that under chronic exposure, the sensitivity of the hepatocytes drastically increased and toxicity of a set of hepatotoxins was detected at clinically relevant concentrations. An interesting example was the chronic toxicity of fialuridine for which hepatotoxicity was mimicked after repeated-dosing in the PHH spheroid model, not possible to detect using previous in vitro systems. Additionally, we provide proof-of-principle that PHH spheroids can reflect liver pathologies such as cholestasis, steatosis and viral hepatitis. Combined, our results demonstrate that the PHH spheroid system presented here constitutes a versatile and promising in vitro system to study liver function, liver diseases, drug targets and long-term DILI.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabinofuranosyluracil / analogs & derivatives
  • Arabinofuranosyluracil / toxicity
  • Cells, Cultured
  • Chemical and Drug Induced Liver Injury / pathology*
  • Chemical and Drug Induced Liver Injury / physiopathology*
  • Hepatocytes / drug effects*
  • Hepatocytes / physiology*
  • Humans
  • Models, Biological
  • Proof of Concept Study
  • Proteome / analysis
  • Spheroids, Cellular / drug effects*
  • Spheroids, Cellular / physiology*


  • Proteome
  • Arabinofuranosyluracil
  • fialuridine