Immunomodulation of activated hepatic stellate cells by mesenchymal stem cells

doi:10.1016/j.bbrc.2007.05.150

Biochemical and Biophysical Research Communications

Volume 363, Issue 2, 16 November 2007, Pages 247-252

https://doi.org/10.1016/j.bbrc.2007.05.150 Get rights and content

Abstract

Bone marrow-derived mesenchymal stem cells (MSCs) have been reported to prevent the development of liver fibrosis in a number of pre-clinical studies. Marked changes in liver histopathology and serological markers of liver function have been observed without a clear understanding of the therapeutic mechanism by which stem cells act. We sought to determine if MSCs could modulate the activity of resident liver cells, specifically hepatic stellate cells (SCs) by paracrine mechanisms using indirect cocultures. Indirect coculture of MSCs and activated SCs led to a significant decrease in collagen deposition and proliferation, while inducing apoptosis of activated SCs. The molecular mechanisms underlying the modulation of SC activity by MSCs were examined. IL-6 secretion from activated SCs induced IL-10 secretion from MSCs, suggesting a dynamic response of MSCs to the SCs in the microenvironment. Blockade of MSC-derived IL-10 and TNF-α abolished the inhibitory effects of MSCs on SC proliferation and collagen synthesis. In addition, release of HGF by MSCs was responsible for the marked induction of apoptosis in SCs as determined by antibody-neutralization studies. These findings demonstrate that MSCs can modulate the function of activated SCs via paracrine mechanisms provide a plausible explanation for the protective role of MSCs in liver inflammation and fibrosis, which may also be relevant to other models of tissue fibrosis.

Section snippets

Materials and methods

Materials were purchased from Sigma–Aldrich, St. Louis, MO unless otherwise stated.

MSC isolation, ex vivo expansion and characterization. Human MSCs were isolated and cultured as previously reported [15]. The surface antigen profile as analyzed by flow cytometry (FACS Calibur, Becton Dickinson) was consistently CD14−, CD34−, CD45−, CD105+, CD106+ and CD44+. Cells were shown to have adipogenic and osteogenic differentiation potential (Suppl. Fig. 1) and were used during passages 4–7.

SC isolation

MSCs inhibit collagen synthesis in activated SCs

The reversal of experimental liver fibrosis in vivo has been highly correlated with a decrease in tissue collagen content [1]. A similar decrease in tissue collagen has been reported after MSC infusion into animals with fibrotic livers [13], [14]. We evaluated whether MSCs can reduce the secretion of procollagen type I C-peptipe (PIP) in SCs. Levels of PIP-secretion by activated SCs (101 ± 11 pg/10⁶cells/day) were significantly reduced at a MSC:SC coculture ratio of 1:10 (41 ± 18 pg/10⁶cells/day; p =

Discussion

Prior studies have shown that transplantation of a CD45− population of bone marrow cells prevented histopathological changes during chronic exposure to hepatotoxins [12], [13], [14]. These observations were correlated with a co-localization of transplanted cells and SCs, a reduction in the number of α-SMA+ cells, decreased tissue collagen content, and increased gelatinase gene expression. These in vivo findings provided a rationale for the therapeutic benefit of MSCs, although evidence

Acknowledgments

We thank Avrum Leeder, Pohun Chris Chen and Luke Selby for the isolation of cells from rat livers. We also acknowledge Robert Crowther’s assistance in cytological and histological preparations and Donald Poulsen’s contributions for medical illustrations. Imaging studies were made possible by the Special Shared Facility in Morphology at the Shriners Hospitals for Children. B.P. is supported by a National Science Foundation predoctoral fellowship. D.V.P. is supported by a fellowship from the

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^☆: Financial support: This work was partially supported by grants from the National Institutes of Health (F32 DK070496, K08 DK66040 and R01 DK43371) and the Shriners Hospitals for Children.

¹: These authors contributed equally to this work.

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Immunomodulation of activated hepatic stellate cells by mesenchymal stem cells☆

Abstract

Section snippets

Materials and methods

MSCs inhibit collagen synthesis in activated SCs

Discussion

Acknowledgments

J. Biol. Chem.

Gastroenterology

Biomaterials

Methods Enzymol.

Am. J. Pathol.

Gastroenterology

Gastroenterology

J. Hepatol.

Combinatorial signaling pathways determine fibroblast proliferation and myofibroblast differentiation

FASEB J.

Reversibility of hepatic fibrosis in experimentally induced cholestasis in rat

Am. J. Pathol.

Mechanisms of spontaneous resolution of rat liver fibrosis. Hepatic stellate cell apoptosis and reduced hepatic expression of metalloproteinase inhibitors

J. Clin. Invest.

The role and regulation of hepatic stellate cell apoptosis in reversal of liver fibrosis

Apoptosis