Working with Immortalized Hepatic Stellate Cell Lines

Friedman, Scott L.; Weiskirchen, Ralf

doi:10.1007/978-1-0716-3207-9_8

Cited by 6 publications

(4 citation statements)

References 28 publications

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“…Therefore, PAV-1 cells are suitable as an alternative HSC in vitro model for various scientific issues in contrast to isolated primary HSCs, which are associated with higher overall costs and always require the use of animals [ 1 , 9 ]. In addition, we showed that PAV-1 lacks the expression of SV40T, which is used to immortalize many other different HSC lines, including LX-1, SV68c-IS, A640-IS (33 °C), IMS/MT(−), IMS/N, HSC-T6, Col-GFP-HSC, JS1 (C57/Bl6 WT), JS2 (C57/Bl6 TLR4 −/− ), and JS3 (C57/Bl6 MyoD88 −/− ) [ 2 , 57 , 58 , 59 , 60 , 61 , 62 , 63 ].…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Immortalized hepatic stellate cell (HSC) lines are useful experimental tools for the study of cellular aspects of hepatic fibrogenesis [ 1 , 2 ]. Like primary HSC, most of these continuously growing cell lines produce a well-developed α-smooth muscle actin (α-SMA) network, express a multitude of characteristic connective tissue markers, and have the capacity to take up and esterify retinol [ 1 , 2 ]. Compared to their primary counterparts that have a limited lifespan, established HSC lines grow faster and can be continuously passaged, indicating that these cells have developed features that allow them to escape from senescence.…”

Section: Introductionmentioning

confidence: 99%

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Genetic Characterization of Rat Hepatic Stellate Cell Line PAV-1

Gäberlein,

Schröder,

Nanda

et al. 2023

Cells

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The rat hepatic stellate cell line PAV-1 was established two decades ago and proposed as a cellular model to study aspects of hepatic retinoic acid metabolism. This cell line exhibits a myofibroblast-like phenotype but also has the ability to store retinyl esters and synthesize retinoic acid from its precursor retinol. Importantly, when cultured with palmitic acid alone or in combination with retinol, the cells switch to a deactivated phenotype in which the proliferation and expression of profibrogenic marker genes are suppressed. Despite these interesting characteristics, the cell line has somehow fallen into oblivion. However, based on the fact that working with in vivo models is becoming increasingly complicated, genetically characterized established cell lines that mimic aspects of hepatic stellate cell biology are of fundamental value for biomedical research. To genetically characterize PAV-1 cells, we performed karyotype analysis using conventional chromosome analysis and multicolor spectral karyotyping (SKY), which allowed us to identify numerical and specific chromosomal alteration in PAV-1 cells. In addition, we used a panel of 31 species-specific allelic variant sites to define a unique short tandem repeat (STR) profile for this cell line and performed bulk mRNA-sequencing, showing that PAV-1 cells express an abundance of genes specific for the proposed myofibroblastic phenotype. Finally, we used Rhodamine-Phalloidin staining and electron microscopy analysis, which showed that PAV-1 cells contain a robust intracellular network of filamentous actin and process typical ultrastructural features of hepatic stellate cells.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genetic Characterization of Rat Hepatic Stellate Cell Line PAV-1

Gäberlein,

Schröder,

Nanda

et al. 2023

Cells

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“…Friedman and Weiskirchen have summarized issues that need to be considered when working with established HSC lines, including experimental recommendations and procedures for media preparation, culture conditions, storage, cryopreservation, banking, and parceling [10]. In addition, information is outlined about safety guidelines, biosafety risk assessment, cell line authentication, and contamination testing.…”

mentioning

confidence: 99%

Book Review: Weiskirchen, R.; Friedman, S.L. Hepatic Stellate Cells: Methods and Protocols, 1st Ed.; Weiskirchen, R., Friedman, S.L., Eds.; Methods in Molecular Biology 2669; Humana Press: New York, NY, USA, 2023; ISBN 978-1-07-163206-2; eISBN: 978-1-0716-3207-9

Weiskirchen

Friedman

2023

Livers

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Cholesterol Depletion Activate Hepatic Stellate Cells Mediated Through SREBP‐2 Signaling

Vijayan,

Perumal

2024

Journal Cellular Physiology

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Liver fibrosis is one of the leading cause of death worldwide. In liver, hepatic stellate cells are the primary cell type that gets activated during fibrosis. LX‐2 cells are human‐derived hepatic stellate cell lines typically employed for studying liver fibrosis mechanisms and screening anti‐fibrotic lead molecules. Although LX‐2 cells are partially activated in culture conditions, numerous stimuli including TGF‐β, H2O2, hypoxia, LPS were reported to activate LX‐2 cells. In this study, for the first time, the effect of cholesterol depletion on LX‐2 cells was studied. Under cholesterol‐depleted conditions, the mRNA and protein expression of HSC activation markers (α‐SMA, GFAP) were significantly increased. Also, the expression of SREBP‐2, HMGCR were significantly upregulated in response to cholesterol depletion. Treatment with fatostatin, a reported SREBP inhibitor abolished nuclear SREBP‐1 and SREBP‐2 expression and regulated the SREBP signaling. Transmission electron microscopic imaging showed distinct ultrastructural changes in response to cholesterol depletion. Furthermore, cholesterol depletion did not affect the cell‐cycle profile of LX‐2 cells compared with untreated while fatostatin treatment induced G2 cell‐cycle arrest. Overall, cholesterol depletion activated LX‐2 cells mediated by SREBP‐2 signaling and therefore could be further employed as stimuli for LX‐2 activation and screening lead molecules targeting SREBPs.

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Working with Immortalized Hepatic Stellate Cell Lines

Cited by 6 publications

References 28 publications

Genetic Characterization of Rat Hepatic Stellate Cell Line PAV-1

Genetic Characterization of Rat Hepatic Stellate Cell Line PAV-1

Book Review: Weiskirchen, R.; Friedman, S.L. Hepatic Stellate Cells: Methods and Protocols, 1st Ed.; Weiskirchen, R., Friedman, S.L., Eds.; Methods in Molecular Biology 2669; Humana Press: New York, NY, USA, 2023; ISBN 978-1-07-163206-2; eISBN: 978-1-0716-3207-9

Cholesterol Depletion Activate Hepatic Stellate Cells Mediated Through SREBP‐2 Signaling

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