Establishment and characterization of primary gallbladder epithelial cell cultures in the prairie dog.
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.
Chapman
W C
WC
Department of Surgery, Vanderbilt University Medical Center, Nashville, Tennessee, 37232, USA.
Fisk
J
J
Schot
D
D
Debelak
J P
JP
Washington
M K
MK
Bluth
R F
RF
Pierce
D
D
Williams
L F
LF
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
D013936:Thymidine