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

W. C. Chapman, J. Fisk, D. Schot, J. P. Debelak, M. K. Washington, R. F. Bluth, D. Pierce, L. F. Williams

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6 Scopus citations

Abstract

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 106 viable cells per animal with viability ranging from 80 to 95%. When plated at 5 x 105 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 antigallbladder 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.

Original languageEnglish
Pages (from-to)35-43
Number of pages9
JournalJournal of Surgical Research
Volume80
Issue number1
DOIs
StatePublished - Nov 1998
Externally publishedYes

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    Chapman, W. C., Fisk, J., Schot, D., Debelak, J. P., Washington, M. K., Bluth, R. F., Pierce, D., & Williams, L. F. (1998). Establishment and characterization of primary gallbladder epithelial cell cultures in the prairie dog. Journal of Surgical Research, 80(1), 35-43. https://doi.org/10.1006/jsre.1998.5401