Human Liver Organoid Culture Protocol

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This protocol outlines a method for culturing normal human liver organoids based on protocols described by Meritxell Huch (1). Starting with a suspension of isolated and dissociated human liver tissue, the reagents and steps necessary to generate and passage human liver organoids are fully detailed. This protocol provides a method for generating an in vitro liver model that can be used for liver regeneration studies, liver disease modeling and toxicology studies, or drug testing. This liver organoid protocol utilizes reagents across the Bio-Techne portfolio including recombinant proteins, small molecules, media supplements, and Cultrex® Organoid Qualified Basement Membrane Extract (BME) as a scaffold.

The protocol includes some special tips and recommendations for culturing human liver organoids including:

  • Recommended media for expanding and differentiating liver organoids
  • Considerations and tips for starting liver organoid cultures
  • Methods to optimize Cultrex Organoid Qualified BME as a matrix scaffold
  • Tips for successful passaging and cryopreservation
  • Data examples for liver organoid characterization


  1. Huch, M. et al. (2015) Cell 160:299.

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