Isolation and long-term culture of gallbladder epithelial cells from wild-type and CF mice.
Mice with targeted disruption of the cftr gene show pathophysiologic changes in the gallbladder, which correlate with hepatobiliary disease seen in cystic fibrosis patients. As gallbladder epithelium secretes mucin, and as this epithelium consists of a relatively homogenous cell type, study of CFTR function in these cells would be beneficial to delineate the complex cellular functions of this protein. The size and anatomic location of the murine gallbladder makes such studies difficult in vivo. Therefore, the need exists for in vitro models of gallbladder epithelium. We describe a method to isolate and culture murine gallbladder epithelium from wild-type and CF mice. Cells were grown in a monolayer on porous inserts over a feeder layer of fibroblasts. These nontransformed cells can be successively passaged and maintain a well-differentiated epithelial cell phenotype as shown by morphologic criteria, characterized by polarized columnar epithelial cells with prominent microvilli and intercellular junctions. Organotypic cultures showed columnar cells simulating in vivo morphology. This culture system should be valuable in delineating cellular processes relating to CFTR in gallbladder epithelium.
Kuver
R
R
Department of Medicine, University of Washington, Seattle, USA.
Savard
C
C
Nguyen
T D
TD
Osborne
W R
WR
Lee
S P
SP
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000032:analysis* / Q000235:genetics; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry / Q000648:ultrastructure; D005704:Gallbladder / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation
D019005:Cystic Fibrosis Transmembrane Conductance Regulator