Isolation and long-term culture of gallbladder epithelial cells from wild-type and CF mice |
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Authors: | Rahul Kuver Christopher Savard Toan D Nguyen William R A Osborne Sum P Lee |
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Institution: | (1) Division of Gastroenterology, University of Washington, 1959 NE Pacific Street, Box 356424, 98195 Seattle, Washington;(2) Department of Medicine, University of Washington and VA Medical Center, 98195 Seattle, Washington;(3) Department of Pediatrics, University of Washington and VA Medical Center, 98195 Seattle, Washington |
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Abstract: | Summary 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. |
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Keywords: | CFTR cell culture gallbladder cystic fibrosis epithelium murine |
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