Human Lung Organoid Culture Protocol

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This protocol outlines a method for culturing normal human lung organoids based on protocols described by Norman Sachs and Hans Clevers (1). The procedure that is described allows for the long-term expansion of airway organoids in vitro from adult human lung tissue. Starting with a suspension of isolated and dissociated human lung tissue, the reagents and steps necessary to generate and passage human lung organoids are fully detailed, allowing for the generation of an in vitro lung model that can be used for pulmonary disease modeling and drug toxicity studies. This lung 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 provides some specific tips and recommendations for culturing human lung organoids including:

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


  1. Sachs, N. et al. (2019) EMBO J. 34:e100300.

Additional Resources

Optimizing Organoid Culture Conditions: The Importance of Growth Factor Bioactivity and Reagent Consistency Application Note

View this application note to learn about the importance of protein bioactivity and reagent consistency for optimizing organoid culture conditions. See how R&D Systems products can help researchers be confident that their culture conditions are consistent and reproducible from one experiment to the next.

Cultrex for Organoid Research

View the different formats of Cultrex Basement Membrane Extract (BME) for 2-D and 3-D cell culture that we offer to support your application-specific needs.

Recombinant Proteins for Organoid Research

Explore our wide selection of proteins for culturing and expanding human lung organoids. From FGFs to Noggin and R-Spondin 1, we have the most trusted, most cited recombinant proteins you need to successfully generate and passage human lung organoids.

Small Molecules for Organoid Research

We offer a wide range of Tocris® small molecules that are commonly used as 3-D growth matrix components for generating different types of organoids and sustaining long-term growth.

Media Supplements for Organoid Research

Our portfolio includes media supplements for use in a variety of cell culture systems. We provide 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 the Building Organoids page on our website for more information on resources and reagents for organoid research including protocols, webinars, additional organoid recipes, organoid or 3-D cell culture-related blogs, scientific posters, literature, and product highlights.