Culture Methods to Study Apical-Specific Interactions using Intestinal Organoid Models

The lining of the gut epithelium is made up a simple layer specialized epithelial cells that expose their apical side to lumen and respond external cues. Recent optimization in vitro culture conditions allows for re-creation intestinal stem cell niche development advanced 3-dimensional (3D) systems recapitulate composition organization epithelium. Intestinal organoids embedded an extracellular matrix (ECM) can be maintained long-term self-organize generate well-defined, polarized encompasses internal exposed basal side. This restrictive nature presents challenges accessing surface limits investigation biological mechanisms such as nutrient uptake host-microbiota/host-pathogen interactions. Here, we describe two methods facilitate access organoid support differentiation specific types. First, show how ECM removal induces inversion polarity generation apical-out 3D organoids. Second, 2-dimensional (2D) monolayers from single suspensions derived organoids, comprised mature differentiated These techniques provide novel tools study apical-specific interactions with cues promote use platform precision medicine.