The effects of Vittel water on the proliferation of fibroblasts, proliferation and differentiation of keratinocytes from human origin were studied. To determine the relative importance of calcium and other elements of the mineral water, cultures were raised in a low-calcium medium (low-Ca medium), in a medium prepared with Vittel water (Vittel medium) and in a medium containing an identical calcium concentration to that of Vittel medium (Ca medium). The fibroblasts and keratinocytes were cultured in immersion for proliferation assays and on a reconstructed epidermis at the air-liquid interface to evaluate keratinocyte differentiation. Vittel medium decreased proliferation of keratinocytes when compared to low-Ca medium. The effect was similar to that of Ca medium at the beginning of the experiment, but significantly higher at day 7. A stratified epithelium appeared with the three types of media when keratinocytes were incubated at the air-liquid interface; however the number of sheets was more regular and greater in Vittel medium and Ca medium than in low-Ca medium. Filaggrin and transglutaminase expression appeared earlier with Vittel medium than with the other media. After 2 weeks, expression of markers was similar in the three media. After 3 weeks culture in Vittel medium, there was a greater expression of filaggrin. Proliferation of young fibroblasts was significantly higher in Vittel medium than in Ca medium. It was lower in low-Ca medium. With old fibroblasts the degree of proliferation was lower than with young fibroblasts. The augmentation of proliferation happen earlier in Vittel medium than in low-Ca medium and Ca medium. Vittel medium regulated the growth rate of old fibroblasts, rendering it identical to that of young fibroblasts in low-Ca medium. The effects of Vittel water were not linked to the sole presence of calcium since, with medium at an equimolecular concentration in calcium, the medium containing Vittel water had a better activity. One explanation of these effects of Vittel water might be the presence of magnesium.