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Isolation and Culture of Primary Murine Hepatic Stellate Cells

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Fibrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1627))

Abstract

Hepatic stellate cells (HSCs) are found in the perisinusoidal space of the liver (i.e., the space of Dissé). They represent 5–8% of the total number of liver cells. In normal liver, these cells have a quiescent phenotype and are characterized by numerous fat vacuoles that store vitamin A in a form of retinyl ester. In injured liver, these cells transdifferentiate into a myofibroblast phenotype, become highly proliferative and are responsible for excess collagen synthesis and deposition during fibrosis. Due to their exceptional pathophysiological relevance, several isolation and purification protocols of primary HSCs have been established that provide the basis for studying HSC biology in vitro. We here describe a method for high-purity isolation of HSCs from mice. This protocol includes the enzymatic digestion of the liver tissue by pronase and collagenase, cellular enrichment by centrifugation of the crude cell suspension through a Nycodenz density gradient, and a final (optional) flow cytometric enrichment that allows generating ultrapure HSC fractions.

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Acknowledgments

This work was supported by grants from the German Research Foundation (SFB/TRR 57 projects P09, P13, and Q3).

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Authors and Affiliations

  1. Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, RWTH University Hospital Aachen, Aachen, Germany

    Sabine Weiskirchen, Carmen G. Tag, Sibille Sauer-Lehnen & Ralf Weiskirchen

  2. Department of Internal Medicine III, RWTH University Hospital Aachen, Aachen, Germany

    Frank Tacke

Authors
  1. Sabine Weiskirchen
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  2. Carmen G. Tag
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  3. Sibille Sauer-Lehnen
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  4. Frank Tacke
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  5. Ralf Weiskirchen
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Corresponding author

Correspondence to Ralf Weiskirchen .

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Editors and Affiliations

  1. Department of Dermatology, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Laure Rittié

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Weiskirchen, S., Tag, C.G., Sauer-Lehnen, S., Tacke, F., Weiskirchen, R. (2017). Isolation and Culture of Primary Murine Hepatic Stellate Cells. In: Rittié, L. (eds) Fibrosis. Methods in Molecular Biology, vol 1627. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7113-8_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7113-8_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7112-1

  • Online ISBN: 978-1-4939-7113-8

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