Primary bile acids (BAs) are produced in the liver from cholesterol and are released into the small intestine to aid in the digestion and absorption of dietary lipids. Most of the BA pool is reabsorbed through enterohepatic circulation, but approximately 5% passes into the colon where the intestinal microbiota convert primary to secondary BAs. These secondary BAs directly impact other members of the microbiome and modulate host metabolism and immunity by signaling through host receptors such as the farnesoid X receptor (FXR). The impact of BAs on barrier immunity in the intestinal epithelium is poorly understood, in part due to a lack of tractable in vitro models. We show that 3-dimensional ‘organotypic’ cultures express bile acid receptors and immune factors, opening the doors to an experimentally tractable model for studying bile acid impacts on immune functions in the intestinal epithelium.