Human Intestinal Organoid Culture Protocol

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This protocol outlines a method for culturing normal human intestinal organoids based on protocols described by C. Pleguezuelos-Manzano and Hans Clevers (1). It is intended to culture organoids from normal human intestinal tissues using Cultrex™ UltiMatrix RGF Basement Membrane Extract as a scaffold, along with recombinant proteins, small molecules, and media supplements available from Bio-Techne. This protocol not only provides you with the recipe for preparing intestinal organoid culture medium, but also notes specific tips used by our in-house cell culture scientists to simplify the culture of human intestinal organoids including:

  • Considerations for starting intestinal organoid cultures from frozen stocks
  • Materials needed for intestinal organoid culture and a detailed recipe for preparing intestinal organoid culture medium
  • Specific instructions for successfully maintaining and passaging human intestinal organoids
  • Image examples of human intestinal organoids cultured from iPSCs or adult stem cells using Cultrex UltiMatrix RGF Basement Membrane Extract and stained for specific intestinal markers
  • Cryobanking and preservation of intestinal organoids for future experiments

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  1. Pleguezuelos-Manzano, C. et al. (2020) Curr. Protoc. Immunol. 130: e106.

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