ESTABLISHMENT AND CHARACTERIZATION OF PRIMARY GALLBLADDER EPITHELIAL-CELL CULTURES IN THE PRAIRIE DOG

Background. The prairie dog has become the established animal gallston e model. This species has a unique propensity to form cholesterol gall stones in response to dietary manipulations. The development of a reli able gallbladder cell culture technique is critical for understanding pathogenic mechanisms of gallstone formation. Materials and meth ods. Prairie dogs underwent laparotomy and cholecystectomy, followed by ini tiation of cell cultures. [H-3]Thymidine incorporation was used to ass ess cell growth, and cell lines were assessed using routine histochemi cal and immunohistochemical staining. Results. Cell yields from prairi e dog gallbladders were 4-8 x 10(6) viable cells per animal with viabi lity ranging from 80 to 95%. When plated at 5 x 10(5) cells/cm(2), cel l clusters, visible within 24 h, coalesced into confluent monolayers w ithin 3-5 days. Cultures remained viable for 6-8 weeks and could be pa ssed for three to four subcultures. Immunohistochemical staining demon strated a high degree of epithelial purity with immunopositivity for A E1/AE3, and cytokeratin, with no vimentin positivity (mesenchymal anti gen). Intracytoplasmic vacuoles demonstrated positive staining for Alc ian blue, periodic acid-Schiff, and mucicarmine and an anti-gallbladde r mucin antibody confirmed the presence of the glycoprotein mucin. Con clusions. This study demonstrates a reliable method for initiation and maintenance of prairie dog gallbladder epithelial cell cultures with a high degree of purity. This technique should allow further studies i nto the pathogenesis of cholesterol gallstones in this model. (C) 1998 Academic Press.