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Testing Cell Migration, Invasion, Proliferation, and Apoptosis in Hepatic Stellate Cells

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Hepatic Stellate Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2669))

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Abstract

The hepatic wound repair process involves cell types including healthy and injured hepatocytes, Kupffer and inflammatory cells, sinusoidal endothelial cells (SECs), and hepatic stellate cells (HSCs). Normally, in their quiescent state, HSCs are a reservoir for vitamin A, but in response to hepatic injury, they become activated myofibroblasts that play a key role in the hepatic fibrotic response. Activated HSCs express extracellular matrix (ECM) proteins, elicit anti-apoptotic responses, and proliferate, migrate, and invade hepatic tissues to protect hepatic lobules from damage. Extended liver injury can lead to fibrosis and cirrhosis, the deposition of ECM that is driven by HSCs. Here we describe in vitro assays that quantify activated HSC responses in the presence of inhibitors targeting hepatic fibrosis.

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References

  1. Subramanian P, Hampe J, Tacke F, Chavakis T (2022) Fibrogenic pathways in metabolic dysfunction associated fatty liver disease (MAFLD). Int J Mol Sci 23(13). https://doi.org/10.3390/ijms23136996

  2. Peiseler M, Schwabe R, Hampe J, Kubes P, Heikenwalder M, Tacke F (2022) Immune mechanisms linking metabolic injury to inflammation and fibrosis in fatty liver disease - novel insights into cellular communication circuits. J Hepatol 77(4):1136–1160. https://doi.org/10.1016/j.jhep.2022.06.012

    Article  CAS  PubMed  Google Scholar 

  3. Tsuchida T, Friedman SL (2017) Mechanisms of hepatic stellate cell activation. Nat Rev Gastroenterol Hepatol 14(7):397–411. https://doi.org/10.1038/nrgastro.2017.38

    Article  CAS  PubMed  Google Scholar 

  4. Singal AG, El-Serag HB (2022) Rational HCC screening approaches for patients with NAFLD. J Hepatol 76(1):195–201. https://doi.org/10.1016/j.jhep.2021.08.028

    Article  PubMed  Google Scholar 

  5. Younossi ZM (2019) Non-alcoholic fatty liver disease – a global public health perspective. J Hepatol 70(3):531–544. https://doi.org/10.1016/j.jhep.2018.10.033

    Article  PubMed  Google Scholar 

  6. Senzolo M, Garcia-Tsao G, Garcia-Pagan JC (2021) Current knowledge and management of portal vein thrombosis in cirrhosis. J Hepatol 75(2):442–453. https://doi.org/10.1016/j.jhep.2021.04.029

    Article  PubMed  Google Scholar 

  7. Xu L, Hui AY, Albanis E, Arthur MJ, O’Byrne SM, Blaner WS et al (2005) Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis. Gut 54(1):142–151. https://doi.org/10.1136/gut.2004.042127

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Borojevic R, Monteiro AN, Vinhas SA, Domont GB, Mourao PA, Emonard H et al (1985) Establishment of a continuous cell line from fibrotic schistosomal granulomas in mice livers. In Vitro Cell Dev Biol 21(7):382–390. https://doi.org/10.1007/BF02623469

    Article  CAS  PubMed  Google Scholar 

  9. Nanda I, Steinlein C, Haaf T, Buhl EM, Grimm DG, Friedman SL et al (2022) Genetic characterization of rat hepatic stellate cell line HSC-T6 for in vitro cell line authentication. Cell 11(11). https://doi.org/10.3390/cells11111783

  10. Friedman SL, Roll FJ (1987) Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation with Stractan. Anal Biochem 161(1):207–218. https://doi.org/10.1016/0003-2697(87)90673-7

    Article  CAS  PubMed  Google Scholar 

  11. Wang J, Leclercq I, Brymora JM, Xu N, Ramezani-Moghadam M, London RM et al (2009) Kupffer cells mediate leptin-induced liver fibrosis. Gastroenterology 137(2):713–723. https://doi.org/10.1053/j.gastro.2009.04.011

    Article  CAS  PubMed  Google Scholar 

  12. Ramezani-Moghadam M, Wang J, Ho V, Iseli TJ, Alzahrani B, Xu A et al (2015) Adiponectin reduces hepatic stellate cell migration by promoting tissue inhibitor of metalloproteinase-1 (TIMP-1) secretion. J Biol Chem 290(9):5533–5542. https://doi.org/10.1074/jbc.M114.598011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Alzahrani B, Iseli T, Ramezani-Moghadam M, Ho V, Wankell M, Sun EJ et al (2018) The role of AdipoR1 and AdipoR2 in liver fibrosis. Biochim Biophys Acta 1864(3):700–708. https://doi.org/10.1016/j.bbadis.2017.12.012

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by a grant from Tropical Australian Academic Health Centre (SF0000121) that is supported by the National Health and Medical Research Council (NHMRC) to LH.

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Correspondence to Lionel Hebbard .

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Wankell, M., Hebbard, L. (2023). Testing Cell Migration, Invasion, Proliferation, and Apoptosis in Hepatic Stellate Cells. In: Weiskirchen, R., Friedman, S.L. (eds) Hepatic Stellate Cells. Methods in Molecular Biology, vol 2669. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3207-9_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3207-9_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3206-2

  • Online ISBN: 978-1-0716-3207-9

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