Long-term culture of cholangiocytes from liver fibro-granulomatous lesions.
BACKGROUND
Extensive bile duct proliferation is a key feature of the tissue reaction to clinical and experimental forms of liver injury. Experimental infection of mice by Schistosoma mansoni is a well-studied model of liver fibrosis with bile duct hyperplasia. However, the regulatory mechanisms of bile duct changes are not well understood. In this study we report the reproducible isolation of long-term cultures of cholangiocytes from mice livers with schistosomal fibrosis.


METHODS
We have isolated a cholangiocyte cell line from Schistosoma-induced liver granulomas using a combination of methods including selective adhesion and isopyknic centrifugation in Percoll.


RESULTS
The cell line was characterized by morphological criteria in optical and transmission electron microscopy, ability to form well differentiated ductular structures in collagen gels and by a positive staining for cytokeratin 18 and cytokeratin 19. To our knowledge, this is the first murine cholangiocyte cell line isolated from schistosomal fibrosis reported in the literature.


CONCLUSIONS
After 9 months and 16 passages this diploid cell line maintained differentiated characteristics and a high proliferative capacity. We believe the method described here may be a valuable tool to study bile duct changes during hepatic injury.
Chiarini
Luciana B
LB
Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21949, Brazil. chiarini@biof.ufrj.br
Takiya
Christina M
CM
Borojevic
Radovan
R
Monteiro
Alvaro N A
AN
D000818:Animals; D001653:Bile Ducts, Intrahepatic / Q000166:cytology* / Q000473:pathology; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D002498:Centrifugation; D003102:Colloids; D003584:Cytological Techniques; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology; D005109:Extracellular Matrix; D005260:Female; D005434:Flow Cytometry; D006099:Granuloma / Q000473:pathology*; D008103:Liver Cirrhosis / Q000382:microbiology / Q000473:pathology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D046529:Microscopy, Electron, Transmission; D011205:Povidone; D012550:Schistosoma mansoni; D012822:Silicon Dioxide
D003102:Colloids; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone