OBJECTIVE To further understand the biology of ureteral cells, we studied the growth characteristics and in vitro cellular interactions of human ureteral uroepithelial cells, smooth muscle cells and myofibroblasts. METHODS The proliferation, morphology and immunohistochemical characteristics of human ureteral cells grown in vitro were evaluated under varying conditions. RESULTS The growth and morphology of ureteral cells were dependent upon media characteristics, especially the calcium concentration and presence of epidermal growth factor and bovine pituitary extract. Cells demonstrated specific stimulatory interactions via both soluble and insoluble factors. Most important, uroepithelial cells and smooth muscle cells displayed reciprocal enhancement of growth. CONCLUSIONS The favorable interactions between ureteral cell types in vitro have implications for future work involving these cells.