Abstract: Mast cells accumulate and persist predominantly in the upper dermis of the skin but the mechanism for this is obscure. The skin is normally exposed to external air, which is essential for the maturation of the epidermis and probably also the dermis. In order to clarify the importance of air exposure on dermal mast cells, skin organ culture at the air–liquid interface (ALI) and submerged (SM) in medium (10% fetal calf serum and Dulbecco's modification of Eagle's medium) was used to study changes in tryptase‐, chymase‐ and Kit‐positive mast cell numbers during cultivation for up to 14 days. In addition, possible apoptosis (TACS TdT in situ apoptosis detection method) in chymase‐positive mast cells was studied during the culture. In the less‐physiologic SM culture, the number of Kit‐positive mast cells decreased rapidly on day 1–2 and tryptase‐positive cells decreased markedly on day 14. This decrease in mast cell numbers can be explained by the finding that a rapid increase in the apoptosis index of mast cells was induced on day 1–2. In contrast, in the more physiologic ALI culture, the number of Kit‐positive cells was sustained over 1–2 days but then decreased on day 7. In addition, tryptase‐positive cells decreased steadily in number but not to the same extent as those in the SM culture. Moreover, the increase in the apoptosis index of mast cells was delayed until day 7 in the ALI culture. Addition of exogenous stem cell factor (up to 200 ng/ml) to the SM culture could not prevent the decay in tryptase‐ and chymase‐positive cells. However, stem cell factor reduced significantly the number of Kit‐positive cells already on day 2 indicating that the cells had responded. Addition of histamine (0.25 or 1 mM) or tumor necrosis factor‐α (500 or 2000 U/ml) caused a decrease in the number of tryptase‐ and Kit‐positive cells in the SM culture. In conclusion, a novel finding was that air exposure in the ALI culture markedly delayed the rapid apoptosis and subsequent decrease in mast cell numbers noted to occur in the SM culture. Stem cell factor could not prevent the rapid decrease in mast cell numbers. Histamine and tumor necrosis factor‐α are possible factors promoting the decline in mast cells.