title: Establishment and characterization of primary gallbladder epithelial cell cultures in the prairie dog.
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.
  
  
  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.
authors.affiliation: Department of Surgery, Vanderbilt University Medical Center, Nashville, Tennessee, 37232, USA.
authors.forename: W C
authors.initials: WC
authors.lastname: Chapman
authors.forename: J
authors.initials: J
authors.lastname: Fisk
authors.forename: D
authors.initials: D
authors.lastname: Schot
authors.forename: J P
authors.initials: JP
authors.lastname: Debelak
authors.forename: M K
authors.initials: MK
authors.lastname: Washington
authors.forename: R F
authors.initials: RF
authors.lastname: Bluth
authors.forename: D
authors.initials: D
authors.lastname: Pierce
authors.forename: L F
authors.initials: LF
authors.lastname: Williams
mesh_terms: D000818:Animals; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D005704:Gallbladder / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D012589:Sciuridae; D013936:Thymidine / Q000378:metabolism
chemical_list: D013936:Thymidine