Pancreatic duct epithelial cells (PDEC) mediate secretion of fluids and electrolytes and are exposed to refluxed bile. In non-transformed cultured dog PDEC, which express many ion transport pathways of PDEC, 1 mM taurodeoxycholic acid (TDCA) stimulated an ¹²⁵I^- efflux inhibited by DIDS and NPPB, and a ⁸⁶Rb⁺ efflux, inhibited by charybdotoxin. Inhibition by BAPTA/AM suggests mediation via increased [Ca²⁺]_i while absence of lactate dehydrogenase release excludes cellular toxicity. At 1 mM, TDCA stimulated a larger ¹²⁵I^- efflux than glycodeoxycholate; two di-hydroxy bile acids, taurochenodeoxycholate and TDCA, were similarly effective while a tri-hydroxy bile acid, taurocholate, was ineffective. In Ussing chambers, 1 mM serosal or 2 mM luminal TDCA stimulated an Isc increase from confluent PDEC monolayers. TDCA also stimulated a) an Isc increase from basolaterally permeabilized PDEC subject to a serosal-to-luminal Cl^- gradient that was inhibited by BAPTA/AM, DIDS, and NPPB, and b) an Isc increase from apically permeabilized PDEC subject to a luminal-to-serosal K⁺ gradient, inhibited by BAPTA/AM and charybdotoxin. Along with the efflux studies, these findings suggest that TDCA interacts directly with PDEC to stimulate Ca²⁺-activated apical Cl^- channels and basolateral K⁺ channels. Monolayer transepithelial resistance was only minimally affected by these concentrations but decreased following exposure to higher TDCA concentrations (2 mM serosal and 4 mM luminal). A secretory role for bile acids should be considered in pancreatic diseases associated with bile reflux.