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BMC Gastroenterology

, 6:13

First Online: 03 April 2006Received: 04 November 2005Accepted: 03 April 2006

Abstract

BackgroundExtensive 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.

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

ResultsThe 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.

ConclusionAfter 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.

AbbreviationsBSSBalanced saline solution

DMEMDulbecco-s minimal essential medium

FBSFetal bovine serum

HEPESN- 2-Hydroxyethyl piperazine-N- 2-ethanesulfonic acid

IPSisotonic Percoll solution

PASperiodic-acid Schiff

PBSPhosphate buffered saline.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-230X-6-13 contains supplementary material, which is available to authorized users.

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Autor: Luciana B Chiarini - Christina M Takiya - Radovan Borojevic - Alvaro NA Monteiro

Fuente: https://link.springer.com/







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