Identification, culture, and characterization of pancreatic stellate cells in rats and humans

doi:10.1016/S0016-5085(98)70209-4

Gastroenterology

Volume 115, Issue 2, August 1998, Pages 421-432

https://doi.org/10.1016/S0016-5085(98)70209-4 Get rights and content

Abstract

Background & Aims: Until now, the basic matrix-producing cell type responsible for pancreas fibrosis has not been identified. In this report, retinoid-containing pancreatic stellate cells (PSCs) in rat and human pancreas are described, and morphological and biochemical similarities to hepatic stellate cells are shown. Methods: Electron and immunofluorescence microscopy (collagen types I and III, fibronectin, laminin, α-actin, and desmin) was performed using pancreatic tissue and cultured PSCs. Extracellular matrix synthesis was shown using quantitative immunoassay and Northern blot analysis. Results: PSCs are located in interlobular areas and in interacinar regions. Early primary cultured PSCs contain retinol and fatty acid retinyl-esters. Addition of retinol to passaged cells resulted in retinol uptake and esterification. During primary culture, the cells changed from a quiescent fat-storing phenotype to a highly synthetic myofibroblast-like cell expressing iso-α-smooth muscle actin (>90%) and desmin (20%–40%) and showing strong positive staining with antibodies to collagen types I and III, fibronectin, and laminin. As determined on protein and messenger RNA level, serum growth factors stimulated the synthesis of collagen type I and fibronectin. Conclusions: The identification of PSCs, particularly in fibrotic areas, and the similarities of these cells to hepatic stellate cells suggest that PSCs participate in the development of pancreas fibrosis.

GASTROENTEROLOGY 1998;115:421-432

Section snippets

Reagents

Materials were purchased from the following sources: collagenase I, rabbit antilaminin, retinol, retinyl-acetate, retinyl-palmitate, yeast transfer RNA, sonicated human placenta DNA, and monoclonal anti–c-fibronectin from Sigma Chemical Co. (Munich, Germany); OptiPrep from Nycomed Pharma (Oslo, Norway); Ficoll and polyU-homopolymer from Pharmacia LKB (Freiburg, Germany); horseradish peroxidase (HRP)–anti-mouse, HRP–anti-rabbit, fluorescein-conjugated streptavidin, biotinylated rabbit anti-mouse

Demonstration of PSCs in the normal and fibrotic pancreas of rats and humans

Cells with cytoplasmic lipid droplets were occasionally visible in the interlobular area or in the interacinar region (Figure 1).

. Transmission electron micrograph showing a PSC in a patient with acute pancreatitis. The PSC is located in close vicinity to a macrophage (Ma) and acinar cells (Ac). Fat droplets (F) and a prominent endoplasmatic reticulum (RER) with cisternae (C) are visible (original magnification 6000×). N, nucleus.

These cells often show a triangular shape, a prominent endoplasmic

Discussion

In this study, retinoid-containing fat-storing cells were identified in the pancreas of rats and humans. The cells contained numerous retinoid-containing fat droplets and were located in the interlobular and interacinar region. Because of their similarity to HSCs, we named them PSCs. The number of the cells increased with age. In addition, these cells are abundant in alcoholic chronic pancreatitis in humans. For further characterization, cells were obtained by outgrowth from small pancreas

Acknowledgements

The authors thank Martina Ehnle, Erika Schmidt, and Adolf Reich for expert technical assistance; Elke Wolff-Hieber for electron- microscopic photographs; and Deedee Falls for proofreading the manuscript.

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^☆: Address requests for reprints to: Max G. Bachem, M.D., Universität Ulm, Klinikum, Institut für Klinische Chemie, Bereichslabor Michelsberg, 89070 Ulm, Germany. e-mail: [email protected]; fax: (49)731-502-6719.

^☆☆: Supported by grant Bausteinförderung P. 347 from the University of Ulm (to M.G.B.) and grant Kn 200/4-1 from the Deutsche Forschungsgemeinschaft (to G.A.).

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Liver, Pancreas, and Biliary TractIdentification, culture, and characterization of pancreatic stellate cells in rats and humans☆,☆☆

Abstract

Section snippets

Reagents

Demonstration of PSCs in the normal and fibrotic pancreas of rats and humans

Discussion

Acknowledgements

Gastroenterology

Cell

Gastroenterology

J Lipid Res

Gastroenterology

J Hepatol

Biochem Biophys Res Commun

Anal Biochem

Exp Cell Res

Hepatology

J Biol Chem

Gastroenterology

Gastroenterology

Cell

Pancreatic extracellular matrix alterations in chronic pancreatitis

Pancreas

Pathology and pathogenesis of diseases of the pancreas

Am J Pathol

Histopathologic and immunohistochemical studies on alcoholic pancreatitis and chronic obstructive pancreatitis: special emphasis on ductal obstruction and genesis of pancreatitis

Am J Gastroenterol

Sequential connective matrix changes in experimental acute pancreatitis. An immunohistochemical and biochemical assessment in rat

Int J Pancreatol

Histopathological and immunohistochemical studies on apparently uninvolved areas of pancreas in patients with acute pancreatitis

Arch Pathol Lab Med

Transforming growth factor β1, extracellular matrix, and inflammatory cells in wound repair using a closed duodenal loop pancreatitis model rat

Scand J Gastroenterol

Accumulation of extracellular matrix and developmental dysregulation in the pancreas by transgenic production of transforming growth factor-β1

Am J Pathol

Chronic pancreatitis

Fibroblast structure and function during regeneration from hormone-induced acute pancreatitis in the rat

Pancreas

Enhancement of transforming growth factor 1 expression in the rat pancreas during regeneration from caerulein-induced pancreatitis

Eur J Clin Invest

Morphological and immunocytochemical identification of periacinar fibroblast-like cells derived from human pancreatic acini

Pancreas

The stellate cell (Ito-cell, fat-storing cell, lipocyte, perisinusoidal cell) of the liver

Virchows Arch B Cell Pathol

Perisinusoidal stellate cells of the liver: important roles in retinol metabolism and fibrosis

FASEB J

Liver, Pancreas, and Biliary Tract
Identification, culture, and characterization of pancreatic stellate cells in rats and humans☆,☆☆