Mechanically dissociated cells from a surgically removed mediastinal C-cell carcinoma (MTC) were cultured over a period of 4 months. The cells of the monolayer culture consisted of clusters of small epithelial-like cells. Using semithin and ultrathin sections, two different types of cells could be characterized by shape of nucleus and by content and distribution of secretory granules. One type of cell showed a more irregularly shaped nucleus, the other contained a large oval nucleus, similar to the normal C-cell of the human thyroid. Calcitonin (CT) and carcinoembryonic antigen (CEA) were measured in supernatants in duplicate by radioimmunoassays. Radioimmuno-detectable CT levels in the supernatant of culture medium varied between 0.8 and 1.6 ng/ml and CEA levels between 5 and 27 ng/ml during a 2-month period. The present study proves that in monolayer-cultured cells of a MTC, metastases continue to produce radioimmuno-detectable CT and CEA. Whether the two different cell types in culture are relevant for carcinoma needs further investigation.