Establishment of an Immortalized Human Hepatic Stellate Cell Line to Develop Antifibrotic Therapies

2003 ◽  
Vol 12 (5) ◽  
pp. 499-507 ◽  
Author(s):  
Norikuni Shibata ◽  
Takamasa Watanabe ◽  
Teru Okitsu ◽  
Masakiyo Sakaguchi ◽  
Michihiko Takesue ◽  
...  
Keyword(s):  
Cell Line ◽  
Hepatic Stellate Cell ◽  
Type I Collagen ◽  
Replicative Senescence ◽  
Stellate Cell ◽  
Smooth Muscle Actin ◽  
Population Doubling ◽  
Type I ◽  
Human Hepatic Stellate Cell ◽  
Ifn Γ

Because human hepatic stellate cells (HSCs) perform a crucial role in the progress of hepatic fibrosis, it is of great value to establish an immortalized human cell line that exhibits HSC characteristics and grows well in tissue cultures for the development of antifibrotic therapies. Thus, we engineered an immortalized human hepatic stellate cell (HSC) line TWNT-4 by retrovirally inducing human telomerase reverse transcriptase (hTERT) into LI 90 cells established from a human liver mesenchymal tumor. Parental LI 90 entered replicative senescence, whereas TWNT-4 showed telomerase activity and proliferated for more than population doubling level (PDL) 200 without any crisis. TWNT-4 expressed platelet-derived growth factor-β receptor (PDGF-βR), α-smooth muscle actin (α-SMA), and type I collagen (α1) and was considered to be an activated form of HSCs. Treatment of TWNT-4 cells with either 100 U/ml of IFN-γ or 1 ng/ml of rapamycin (Rapa) for 14 days led to lower expression of type I collagen (α1) at RNA and protein levels. Exposure of TWNT-4 cells to both of IFN-γ (10 U/ml) and Rapa (0.1 ng/ml) for 14 days effectively decreased the expression of type I collagen (α1), PDGF-βR, and α-SMA expression and suppressed TGF-β1 secretion of TWNT-4 cells. We successfully induced apoptosis by transducing TNF-related apoptosis-inducing ligand (TRAIL) into TWNT-4 cells using adenovirus vectors Ad/GT-TRAIL and Ad/PGK-GV-17. These findings suggested that immortalized activated HSC line TWNT-4 would be a useful means to develop antifibrotic therapies.

NP603, a novel and potent inhibitor of FGFR1 tyrosine kinase, inhibits hepatic stellate cell proliferation and ameliorates hepatic fibrosis in rats

2011 ◽  
Vol 301 (2) ◽  
pp. C469-C477 ◽  
Author(s):  
Nan Lin ◽  
Si Chen ◽  
Weidong Pan ◽  
Linan Xu ◽  
Kunpeng Hu ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Hepatic Fibrosis ◽  
Hepatic Stellate Cell ◽  
Type I Collagen ◽  
Kinase Inhibitor ◽  
Stellate Cell ◽  
Smooth Muscle Actin ◽  
Type I ◽  
Promising Therapeutic Approach ◽  
The Impact

Fibroblast growth factor 2 (FGF-2) and its main receptor FGFR1 have been shown to promote hepatic stellate cell (HSC) activation and proliferation. However, scant information is available on the anti-fibrogenic activity of FGFR1 inhibitors. The aim of this study was to assess the impact of a selective FGFR1 tyrosine kinase inhibitor NP603 on HSC proliferation and hepatic fibrosis. We demonstrated that rat primary HSCs secreted significant amounts of FGF-2, and its tyrosine phosphorylation of FGFR1 was attenuated by NP603. NP603 inhibited HSC activaton by measuring the expression of α-smooth muscle actin (α-SMA) and the production of type I collagen using ELISA. Furthermore, NP603 (25 μM) in vitro strongly suppressed HSC growth induced by FGF-2 (10 ng/ml) and FCS. This effect correlated with the suppression of extracellular-regulated kinase (ERK) activity and its downstream targets cyclin D1 and p21. In addition, PO NP603 (20 mg·kg−1·day−1) administration significantly decreased hepatic collagen deposition and α-SMA expression in CCl4-treated rats. Collectively, these studies suggest that selective blocking of the FGFR1-mediated pathway could be a promising therapeutic approach for the treatment of hepatic fibrosis.

scholarly journals Dietary Flavonoid Hyperoside Induces Apoptosis of Activated Human LX-2 Hepatic Stellate Cell by Suppressing Canonical NF-κB Signaling

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Liwen Wang ◽  
Zhiwei Yue ◽  
Mengzheng Guo ◽  
Lianying Fang ◽  
Liang Bai ◽  
...  
Keyword(s):  
Hepatic Stellate Cell ◽  
Type I Collagen ◽  
Stellate Cell ◽  
Smooth Muscle Actin ◽  
Binding Activity ◽  
Type I ◽  
Altered Expression ◽  
Dna Binding Activity ◽  
Dietary Flavonoid ◽  
Induced Apoptosis

Hyperoside, an active compound found in plants of the generaHypericumandCrataegus, is reported to exhibit antioxidant, anticancer, and anti-inflammatory activities. Induction of hepatic stellate cell (HSC) apoptosis is recognized as a promising strategy for attenuation of hepatic fibrosis. In this study, we investigated whether hyperoside treatment can exert antifibrotic effects in human LX-2 hepatic stellate cells. We found that hyperoside induced apoptosis in LX-2 cells and decreased levels ofα-smooth muscle actin (α-SMA), type I collagen, and intracellular reactive oxygen species (ROS). Remarkably, hyperoside also inhibited the DNA-binding activity of the transcription factor NF-κB and altered expression levels of NF-κB-regulated genes related to apoptosis, including proapoptotic genesBcl-Xs,DR4,Fas, andFasLand anti-apoptotic genesA20,c-IAP1,Bcl-XL, andRIP1. Our results suggest that hyperoside may have potential as a therapeutic agent for the treatment of liver fibrosis.

Growth Arrest and Decrease of α-SMA and Type I Collagen Expression by Palmitic Acid in the Rat Hepatic Stellate Cell Line PAV-1

2006 ◽  
Vol 51 (5) ◽  
pp. 986-995 ◽  
Author(s):  
Armand Abergel ◽  
Vincent Sapin ◽  
Nicolas Dif ◽  
Christophe Chassard ◽  
Claude Darcha ◽  
...  
Keyword(s):  
Palmitic Acid ◽  
Cell Line ◽  
Hepatic Stellate Cell ◽  
Type I Collagen ◽  
Stellate Cell ◽  
Growth Arrest ◽  
Type I ◽  
Collagen Expression

scholarly journals Establishment and Characterization of a Novel Fibroblastic Cell Line (SCI13D) Derived from the Broncho-Alveolar Lavage of a Patient with Fibrotic Hypersensitivity Pneumonitis

Biomedicines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1193
Author(s):  
Paolo Giannoni ◽  
Marco Grosso ◽  
Giuseppina Fugazza ◽  
Mario Nizzari ◽  
Maria Cristina Capra ◽  
...  
Keyword(s):  
Cell Line ◽  
Hypersensitivity Pneumonitis ◽  
Type I Collagen ◽  
Smooth Muscle Actin ◽  
Usual Interstitial Pneumonia ◽  
Type I ◽  
Fibroblastic Cell ◽  
Primary Fibroblasts

Hypersensitivity pneumonitis (HP) is a diffuse interstitial lung disease (ILD) caused by the inhalation of a variety of antigens in susceptible individuals. Patients with fibrotic HP (fHP) may show histopathological and radiological manifestations similar to patients with idiopathic pulmonary fibrosis (usual interstitial pneumonia-like pattern of fibrosis) that are associated with a worse prognosis. We describe here the establishment and characterization of a fibroblastic cell line derived from the broncho-alveolar lavage (BAL) of a patient with fHP, a 53 year old man who presented at our Pneumology Unit with cough and dyspnea. The fHP diagnosis was based on international criteria and multidisciplinary discussion. Primary fibroblasts were expanded in vitro until passage 36. These fibroblasts displayed morpho/phenotypical features of myofibroblasts, showing high positivity for α-smooth muscle actin, type I collagen, and fibronectin as determined by quantitative RT-PCR and cyto-fluorographic analysis. Cytogenetic analyses further evidenced trisomy of chromosome 10, which interestingly harbors the FGF2R gene. To our knowledge, this is the first fibroblastic cell line derived from an fHP patient and might, therefore, represent a suitable tool to model the disease in vitro. We preliminarily assessed here the activity of pirfenidone, further demonstrating a consistent inhibition of cells growth by this antifibrotic drug.

Human hepatic stellate cell line (LX-2) exhibits characteristics of bone marrow-derived mesenchymal stem cells

2011 ◽  
Vol 91 (3) ◽  
pp. 664-672 ◽  
Author(s):  
Andrielle Castilho-Fernandes ◽  
Danilo Candido de Almeida ◽  
Aparecida Maria Fontes ◽  
Fernanda Ursoli Ferreira Melo ◽  
Virgínia Picanço-Castro ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cell Line ◽  
Hepatic Stellate Cell ◽  
Stellate Cell ◽  
Human Hepatic Stellate Cell
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