Bile secretion by liver parenchymal cells is the result of vectorial transcellular transport of solutes and involves the coordinated action of transport proteins at the basolateral (sinusoidal) and apical (canalicular) membranes of the hepatocyte. A complex network of signals controls uptake and efflux transporters on a long- and a short-term timescale, including regulation at the level of gene transcription, protein translation and maturation, covalent modification, and dynamic localization of transporter proteins, as well as substrate availability. Evidence has shown that the hepatocellular hydration state exerts powerful control on the transcellular transport of solutes, such as conjugated bile acids and glucuronide and glutathione conjugates. This is of physiological significance because liver cell hydration is a dynamic parameter, which changes within minutes under the influence of hormones, nutrients, and oxidative stress. Thus, osmoregulation of bile formation is of physiological and pathophysiological interest.