A procedure is described for the tissue culture of epithelial cells from Barrett's specialized columnar epithelium. The method employs use of nonenzymatic disaggregation techniques and use of a specialized growth medium, M-19. Successful growth can be achieved in 60-70% of cases. Characteristics of four cultures are presented including PAS, Alcian blue, and cytokeratin staining properties. Ultrastructural studies showed the presence of distended rough endoplasmic reticulum, abundant Golgi apparatus, glycogen granules, and cell-cell junctional complexes. Colony formation in anchorage-independent growth in soft agar was not observed in any culture. The cultured cells were not capable of indefinite growth. Thus, they do not behave as fully transformed cells. Availability of this culture system should be of use to the study of the biology of this metaplastic premalignant lesion.