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Vinculin and cellular retinol-binding protein-1 are markers for quiescent and activated hepatic stellate cells in formalin-fixed paraffin embedded human liver

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Abstract

Hepatic stellate cells (HSCs) have important roles in the pathogenesis of liver fibrosis and cirrhosis. As response to chronic injury HSCs are activated and change from quiescent into myofibroblast-like cells. Several HSC-specific markers have been described in rat or mouse models. The aim of our work was to identify the best marker(s) for human HSCs. To this end we used the automated high throughput NexES IHC staining device (Ventana Medical Systems) to incubate sections under standardized conditions. Formalin fixed paraffin embedded (FFPE) normal and diseased human livers were studied. With immunohistochemistry we examined the expression of synemin, desmin, vimentin, vinculin, neurotrophin-3 (NT-3), α-smooth muscle actin (α-SMA), cellular retinol-binding protein-1 (CRBP-1), glial fibrillary acidic protein (GFAP), cysteine- and glycine-rich protein 2 (CRP2), and cytoglobin/stellate cell activation-associated protein (cygb/STAP). This is the first study in which a series of HSC markers is compared on serial FFPE human tissues. CRBP-1 clearly stains lobular HSCs without reacting with smooth muscle cells (SMCs) and shows variable cholangiocyte positivity. Vinculin has a similar staining pattern as CRBP-1 but additionally stains SMCs, and (myo)fibroblasts. In conclusion, we therefore propose to use CRBP-1 and/or vinculin to stain HSCs in human liver tissues.

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Abbreviations

HSCs:

Hepatic stellate cells

FFPE:

Formalin fixed paraffin embedded

NT-3:

Neurotrophin-3

α-SMA:

α-Smooth muscle actin

CRBP-1:

Cellular retinol-binding protein-1

GFAP:

Glial fibrillary acidic protein

CRP2:

Cysteine- and glycine-rich protein 2

cygb/STAP:

Cytoglobin/stellate cell activation-associated protein

SMCs:

Smooth muscle cells

ECM:

Extracellular matrix

PBS:

Phosphate buffered saline

DAB:

3, 3′-Diaminobenzidine

AEC:

3-Amino-9-ethylcarbazole

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Acknowledgments

The authors express their warmest thanks to Christel Van Den Broeck for performing the immunohistochemical stainings, Prof. Weiskirchen and Kawada for the gift of CRP2 and cygb/STAP antibodies. This work was supported by FWO-V (Fonds voor Wetenschappelijk Onderzoek – Vlaanderen), grants G.0512.04, G.0652.06, G.0229.08, G.0651.06; OZR (Research Council of Vrije Universiteit Brussel) grants OZR1432BOF, OZR1149BOF, GOA48, OZR1600BOF, OZR1226, OZR1219; EASL (European Association for the Study of the Liver) 2007 Sheila Sherlock Entry Level Fellowship; I.A.P. (Belgian Science Policy Bureau) phase VI contract P6/36; IWT/SB/61314; EU 6th Framework program. Roche and Schering-Plough unconditional grant.

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Authors and Affiliations

  1. Department of Cell Biology, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel, Jette, Belgium

    Elke Van Rossen, Leo Adrianus van Grunsven, Hendrik Reynaert & Albert Geerts

  2. Laboratory of Morphology and Molecular Pathology, University of Leuven, Leuven, Belgium

    Sara Vander Borght & Tania Roskams

  3. Division Livestock-Nutrition-Quality, Department of Biosystems, Katholieke Universiteit Leuven, Leuven, Belgium

    Veerle Bruggeman

  4. Department of Nutrition, Faculty of Medicine, University of Oslo, Oslo, Norway

    Rune Blomhoff

  5. Laboratory for Cell Biology, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, 1090, Brussel-Jette, Belgium

    Elke Van Rossen

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  1. Elke Van Rossen
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  2. Sara Vander Borght
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  3. Leo Adrianus van Grunsven
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Correspondence to Elke Van Rossen.

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E. Van Rossen, S. Vander Borght, T. Roskams, and A. Geerts contributed equally to this manuscript.

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Van Rossen, E., Vander Borght, S., van Grunsven, L.A. et al. Vinculin and cellular retinol-binding protein-1 are markers for quiescent and activated hepatic stellate cells in formalin-fixed paraffin embedded human liver. Histochem Cell Biol 131, 313–325 (2009). https://doi.org/10.1007/s00418-008-0544-2

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