Human Small Intestinal Organoids Differentiation Protocols Towards All Mature Epithelial Lineages

The small intestine plays a crucial role in nutrient uptake and metabolism regulation while maintaining a barrier that protects against the harsh environment of the lumen. These different tasks are carried out by a multitude of cell types, continuously generated by the resident stem cell population, and whose specification is under tight regulatory control. Organoids are self-organizing 3D structures derived from tissue-resident stem cells, which retain the ability to differentiate into all other epithelial lineages. Protocols to enrich for the various cell subtypes in the intestine have greatly advanced in recent years. Here, we provide an updated overview of these protocols, which direct human small intestinal organoids differentiation into various mature cell lineages, recapitulating the in vivo gut.