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
References
- Huch, M. et al. (2015) Cell 160:299.
Additional Resources
Optimizing Organoid Culture Conditions: The Importance of Growth Factor Bioactivity and Reagent Consistency Application Note
View this application note to learn how important growth factor bioactivity and reagent consistency are for optimizing organoid culture and why R&D Systems proteins for organoid research are better than proteins provided by our leading competitors.
Cultrex for Organoid Research
Explore the different types of Cultrex Basement Membrane Extracts (BME), Cultrex Extracellular Matrix proteins, and Cultrex BME Cell Invasion and Migration Assays that we offer .
Recombinant Proteins for Organoid Research
View our wide range of proteins for culturing and expanding human liver organoids. From EGF, FGF-10, and HGF to Wnt-3a and R-Spondin 1, we offer the highest quality bioactive recombinant proteins to successfully generate and passage human liver organoids.
Small Molecules for Organoid Research
Our portfolio includes a large selection of Tocris® small molecules that are used as growth matrix components for the generation of different types of organoids and the maintenance of long-term growth.
Choosing the Right Model System for Hepatotoxicity Scientific Poster
Use the data in this scientific poster to learn about the advantages of evaluating liver toxicity with three different in vitro hepatotoxicity model systems: induced pluripotent stem cell (iPSC)-derived hepatocytes, 3-D liver organoids, and HepG2 spheroids.
Media Supplements for Organoid Research
If you are in need of media supplements for organoid research, we offer general use media supplements, supplements for specialized cell culture, and protein-free media for the cryopreservation of cells.
All Resources and Reagents for Organoid Research
Explore our complete collection of resources and reagents for organoid research including organoid recipes, protocols, webinars, organoid-related blogs, scientific posters, literature, and product highlights.