Development and functional characterization of extrahepatic cholangiocyte lines from normal rats

Venter, Julie; Francis, Heather; Meng, Fanyin; DeMorrow, Sharon; Kennedy, Lindsey; Standeford, Holly; Hargrove, Laura; Wu, Nan; Wan, Ying; Frampton, Gabriel; McMillin, Matthew; Marzioni, Marco; Gaudio, Eugenio; Onori, Paolo; Glaser, Shannon; Alpini, Gianfranco

doi:10.1016/j.dld.2015.07.012

Cited by 11 publications

(6 citation statements)

References 53 publications

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“…In contrast, most of conventional BEC markers, such as CK19 (Krt19), Epcam, Cftr, and anion exchanger protein 2, were equally expressed between Trop2 À and Trop2 þ BECs (Supplemental Figure S3), consistent with the immunohistological data of mouse EHBD (Figure 1, A and C) and rat EHBD. 24 These results supported the hypothesis that Trop2 is a valuable cell surface marker to separate the luminal epithelium and PBG of EHBD (ie, LBECs and PBECs).…”

Section: Trop2 Is Predominantly Expressed In Lbecs But Not In Pbecssupporting

confidence: 78%

Characterization of Peribiliary Gland–Constituting Cells Based on Differential Expression of Trophoblast Cell Surface Protein 2 in Biliary Tract

Matsui

Harada

Miyata

et al. 2018

The American Journal of Pathology

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Peribiliary glands (PBGs) are accessory glands with mucinous and serous acini in the biliary tree. The PBG is composed of a heterogeneous cell population, such as mucus- and pancreatic enzyme-producing epithelial cells, whereas it constitutes niches for multipotential stem/progenitor cells in the human extrahepatic bile duct (EHBD). By contrast, the nature of PBGs in the mouse EHBD remains unclear. Our aim was to establish a method for isolating and characterizing PBG-constituting cells in the mouse EHBD. We found that trophoblast cell surface protein 2 (Trop2) was expressed in the luminal epithelium of mouse EHBD exclusively, but not in the PBG. On the basis of the differential expression profile of Trop2, lumen-forming biliary epithelial cells (LBECs) and PBG-constituting epithelial cells (PBECs) were separately isolated for further characterization. Gene expression analysis revealed that the isolated mouse PBECs expressed several marker genes related to human PBGs. In the colony formation assay, PBECs showed significantly higher colony formation capacity than LBECs. In the organoid formation assay, PBECs formed cystic organoid with LBEC-like phenotype. Interestingly, PBECs proliferated, accompanied by reexpression of Trop2 in vivo after bile duct ligation. Furthermore, the unique expression profile of Trop2 was conserved in human EHBD. Our findings indicate that Trop2 is a useful marker in investigating the pathophysiological roles and characteristics of mouse and human PBGs in biliary diseases.

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Section: Trop2 Is Predominantly Expressed In Lbecs But Not In Pbecssupporting

confidence: 78%

Characterization of Peribiliary Gland–Constituting Cells Based on Differential Expression of Trophoblast Cell Surface Protein 2 in Biliary Tract

Matsui

Harada

Miyata

et al. 2018

The American Journal of Pathology

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“…The epithelial injury in experimental BA has been mechanistically linked to innate immunity 12, 25–27 . Here, we extend the innate signals to NLRP3 and IL-1R1, individual elements of a molecular complex responsive to danger signals from infection, stress or sterile damage.…”

Section: Discussionmentioning

confidence: 99%

Regulation of epithelial injury and bile duct obstruction by NLRP3, IL-1R1 in experimental biliary atresia

Yang

Mizuochi

Shivakumar

et al. 2018

Journal of Hepatology

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Biliary atresia is a severe inflammatory and obstructive disease of bile ducts occurring in infancy. Although the cause is unknown, activation of the innate and adaptive immune systems injures the bile duct epithelium. In this study we found that patients' livers had increased expression of inflammasome genes. Using mice engineered to inactivate individual inflammasome genes, the epithelial injury and bile duct obstruction were prevented by the loss of Il1r1 or Nlrp3, with a decreased activation of natural killer cells and expression of cytokines and chemokines. In contrast, the loss of Casp1 did not change the disease phenotype. Combined, the findings point to a differential role of inflammasome gene products in the pathogenic mechanisms of biliary atresia.

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“…Antibodies for immunohistochemistry and immunofluorescence were obtained from Abcam (Cambridge, MA) unless indicated otherwise. Total RNA was isolated from total liver tissues, purified cholangiocytes, and selected cell lines using the TRI Reagent from Sigma Life Science and reverse transcribed with the Reaction Ready First Strand cDNA synthesis kit (SABiosciences, Frederick, MD) as described 27 . Total RNA was extracted from HSCs (isolated by Laser Capture Microdissection, LMD) using the Arcturus ® PicroPure ® RNA Isolation Kit (Applied Biosystems; Waltham, MA) according to the manufacturers protocol.…”

Section: Methodsmentioning

confidence: 99%

Knockout of microRNA-21 reduces biliary hyperplasia and liver fibrosis in cholestatic bile duct ligated mice

Kennedy

Meng

Venter

et al. 2016

Laboratory Investigation

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Cholestasis is a condition that leads to chronic hepatobiliary inflammation, fibrosis, and eventually cirrhosis. Many microRNAs (miRs) are known to play a role in fibrosis progression; however, the role of miR-21 during cholestasis remains unknown. Therefore, the aim of this study was to elucidate the role of miR-21 during cholestasis-induced biliary hyperplasia and hepatic fibrosis. Wild-type (WT) and miR21−/− mice underwent sham or bile duct ligation (BDL) for 1 wk, before evaluating liver histology, biliary proliferation, hepatic stellate cell (HSC) activation, fibrotic response, and Smad-7 expression. In vitro, immortalized murine biliary cell lines (IMCL) and human hepatic stellate cell line (hHSC) were treated with either miR-21 inhibitor or control before analyzing proliferation, apoptosis, and fibrotic responses. In vivo, the levels of miR-21 were increased in total liver and cholangiocytes after BDL, and loss of miR-21 decreased the amount of BDL-induced biliary proliferation and intrahepatic biliary mass. Also, loss of miR-21 decreased BDL-induced HSC activation, collagen deposition, and expression of the fibrotic markers TGF-β1 and α-SMA. In vitro, IMCL and hHSCs treated with miR-21 inhibitor displayed decreased proliferation and expression of fibrotic markers and enhanced apoptosis when compared to control treated cells. Furthermore, mice lacking miR-21 show increased Smad-7 expression, which may be driving the decrease in biliary hyperplasia and hepatic fibrosis. During cholestatic injury miR-21 is increased and leads to increased biliary proliferation and hepatic fibrosis. Local modulation of miR-21 may be a therapeutic option for patients with cholestasis.

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Development and functional characterization of extrahepatic cholangiocyte lines from normal rats

Cited by 11 publications

References 53 publications

Characterization of Peribiliary Gland–Constituting Cells Based on Differential Expression of Trophoblast Cell Surface Protein 2 in Biliary Tract

Characterization of Peribiliary Gland–Constituting Cells Based on Differential Expression of Trophoblast Cell Surface Protein 2 in Biliary Tract

Regulation of epithelial injury and bile duct obstruction by NLRP3, IL-1R1 in experimental biliary atresia

Knockout of microRNA-21 reduces biliary hyperplasia and liver fibrosis in cholestatic bile duct ligated mice

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