Hepatocytes, which were Mg2+-depleted during isolation, took up Mg2+ during reincubation. Mg2+ uptake was dependent on the concentration of extracellular Mg2+, Na+, Cl-, bicarbonate and phosphate. Li+ and choline+ did not substitute for extracellular Na+ in Mg2+ influx. Mg2+ influx was maximal when all three anion species were present, and did not occur when these anions were replaced by gluconate. Bicarbonate, phosphate and Cl- could substitute for each other. Mg2+ uptake in hepatocytes was inhibited by p-chloromercuribenzene sulfonate, ouabain, gramicidin D, amiloride and verapamil. The results were explained by the assumption that net Mg2+ influx in hepatocytes is operating via electroneutral Na+, Mg2+/anion cotransport driven by the Na+ gradient. However, electrogenic Mg2+ uptake gated by extracellular Na+ and anions could not be excluded.