ORGANOTYPIC CULTURE OF HUMAN GALLBLADDER EPITHELIUM

We have modified methods of growing human gallbladder epithelial cells in monolayer and organotypic culture. These cells were grown in the p resence of fetal bovine serum and with coculture of feeder layers of h uman gallbladder fibroblasts. Human gallbladders were obtained from ch olecystectomy specimens, and the cells were dissociated with trypsin/E DTA. Cells, which were grown with feeder layer on collagen-coated plat es in the presence of 10% FBS, grew rapidly and formed islands uf cubo idal cells with morphology typical of epithelial cells in culture. The y could be passaged up to four times. The cells were also successfully grown by organotypic technique producing a monolayer of tall, columna r, palisade, epithelial cells. These cells, both in monolayer and in o rganotypic culture, were positive to antibodies for simple epithelial keratin and negative to antibody for vimentin or any of the mesenchyma l antibodies. These cells respond to agonists (prostaglandin E(2), iso proterenol) by the intracellular generation of cAMP. Secreted mucin on the apical surface stained strongly with periodic acid-Schiff. Organo typic culture of human gallbladder epithelium may serve as a cell prep aration for the study of pathobiology of columnar epithelial cells. (C ) 1995 Academic Press, Inc.