The appearance of large human basophilic cells containing histamine was followed to reflect cell maturation in cultures of peripheral blood leukocytes. Aliquots were taken on day 0 and after 7, 9-10 and 14 days in culture, and the basophilic cell numbers were determined in 200 microliter cytospin specimens stained with Wright's stain. The number of small basophils initially present declined during the culture period. The number of spontaneously appearing large basophilic cells varied among the cultures from different individuals and was slightly, though not significantly, higher in atopic individuals than in normals. In cultures of cells from atopic individuals, the number of large basophilic cells was reproducibly increased by the addition of appropriate allergen to the cultures. In cultures of cells from normal individuals, the number was increased two- to six-fold by adding conditioned medium (CM), either from 3-day cultures of cells from atopic individuals stimulated with allergen or with CM from the Mo T cell leukemic line. The addition of mitomycin C decreased the number of large basophilic cells and cycloheximide abrogated their appearance.