Establishment and characterization of primary gallbladder epithelial cell cultures in the prairie dog.

Chapman WC, Fisk J, Schot D, Debelak JP, Washington MK, Bluth RF, Pierce D, Williams LF
J Surg Res. 1998 80 (1): 35-43

PMID: 9790812 · DOI:10.1006/jsre.1998.5401

BACKGROUND - The prairie dog has become the established animal gallstone model. This species has a unique propensity to form cholesterol gallstones in response to dietary manipulations. The development of a reliable gallbladder cell culture technique is critical for understanding pathogenic mechanisms of gallstone formation.

MATERIALS AND METHODS - Prairie dogs underwent laparotomy and cholecystectomy, followed by initiation of cell cultures. [3H]Thymidine incorporation was used to assess cell growth, and cell lines were assessed using routine histochemical and immunohistochemical staining.

RESULTS - Cell yields from prairie dog gallbladders were 4-8 x 10(6) viable cells per animal with viability ranging from 80 to 95%. When plated at 5 x 10(5) cells/cm2, cell clusters, visible within 24 h, coalesced into confluent monolayers within 3-5 days. Cultures remained viable for 6-8 weeks and could be passed for three to four subcultures. Immunohistochemical staining demonstrated a high degree of epithelial purity with immunopositivity for AE1/AE3, and cytokeratin, with no vimentin positivity (mesenchymal antigen). Intracytoplasmic vacuoles demonstrated positive staining for Alcian blue, periodic acid-Schiff, and mucicarmine and an anti-gallbladder mucin antibody confirmed the presence of the glycoprotein mucin.

CONCLUSIONS - 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 into the pathogenesis of cholesterol gallstones in this model.

Copyright 1998 Academic Press.

MeSH Terms (9)

Animals Cell Division Cells, Cultured Epithelial Cells Gallbladder Immunohistochemistry Microscopy, Electron Sciuridae Thymidine

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