Calcium is one of the most important substances affecting the life of molluscs, and vitamin D was shown to be an essential nutrient for land snails. In an attempt to elucidate the role that vitamin D plays in calcium metabolism of land snails, we have developed a procedure for the isolation of specialized calcium cells from digestive gland of land snails, and were able to culture these cells. The effect of vitamin D metabolites on the intracellular exchangeable calcium and alkaline phosphatase activity was studied. The metabolites tested were 25-hydroxycholecalciferol (25(OH)D3), 24,25-dihydroxycholecalciferol (24,25(OH)2D3), 1,25-dihydroxycholecalciferol (1,25(OH)2D3), and the molluscan metabolite E. 25(OH)D3 was found to be the most active sterol in elevating intracellular exchangeable calcium and the activity of alkaline phosphatase, and the molluscan metabolite E was found to be the most potent sterol in the suppression of alkaline phosphatase activity. 1,25(OH)2D3 was shown to suppress both activities at high concentrations, and 24,25(OH)2D3 increased the intracellular exchangeable calcium only at high concentrations. Thus, 25(OH)D3 which is regarded as a storage form of vitamin D and devoid of biological activity, seems to be biologically active in invertebrates.