FIGURE 6.

Functional amino acid transport assay using intestinal organoid monolayers. (A) 3-Dimensional organoids were dissociated into single cells and plated on Matrigel precoated (0.5% v/v) Transwell inserts. The monolayers were grown to confluence for 1 or 4 days and generated TEER values >750 Ω/cm2. The apical medium was replaced with DMEM and basolateral medium with HBSS. Amino acids (AA) and other molecules were measured in the basal compartment using TQMS. (B) AA concentrations (Log values) of individual amino acids and other molecules for 4 replicate Transwell inserts of ileum organoids. (C) AA concentrations in the basolateral compartment show increased transport of essential AA’s (orange).