Establishment of a novel in vitro co-culture system of enteric neurons and Caco-2 cells for evaluating the effect of enteric nervous system on transepithelial transport of drugs

[Display omitted] •Isolated NCS-like cells from LMMP were proliferated and differentiated into neurons.•Establishment of a novel co-culture system of Caco-2 cells and enteric neurons.•Enteric neurons mainly affected the passive transport via paracellular pathway. The gastrointestinal tract is innervated by extrinsic autonomic nerves and intrinsic enteric nervous system (ENS). However, the role of ENS in drug absorption has remained to be clarified. To investigate the effect of ENS on drug transport across the intestinal epithelial cells, we established a novel co-culture system of Caco-2 cells and enteric neurons differentiated from neural crest stem (NCS)-like cells isolated from mouse longitudinal muscle/myenteric plexus (LMMP). Immunostaining analysis revealed that the proportions of neuron, glia, and NCS-like cells were only