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Fibrogenic potential of human multipotent mesenchymal stromal cells in injured liver.

Baertschiger RM, Serre-Beinier V, Morel P, Bosco D, Peyrou M, Clément S, Sgroi A, Kaelin A, Buhler LH, Gonelle-Gispert C - PLoS ONE (2009)

Bottom Line: Expansion capacity of pediatric MSC was significantly higher when compared to adult MSC.In conclusion, when transplanted into an injured or regenerating liver, MSC differentiated into myofibroblasts with development of fibrous tissue, regardless of donor age.These results indicate that MSC in certain circumstances might be harmful due to their fibrogenic potential and this should be considered before potential use of MSC for cell therapy.

View Article: PubMed Central - PubMed

Affiliation: Surgical Research Unit, Department of Surgery, University Hospital Geneva, Geneva, Switzerland.

ABSTRACT
Multipotent mesenchymal stromal cells (MSC) are currently investigated clinically as cellular therapy for a variety of diseases. Differentiation of MSC toward endodermal lineages, including hepatocytes and their therapeutic effect on fibrosis has been described but remains controversial. Recent evidence attributed a fibrotic potential to MSC. As differentiation potential might be dependent of donor age, we studied MSC derived from adult and pediatric human bone marrow and their potential to differentiate into hepatocytes or myofibroblasts in vitro and in vivo. Following characterization, expanded adult and pediatric MSC were co-cultured with a human hepatoma cell line, Huh-7, in a hepatogenic differentiation medium containing Hepatocyte growth factor, Fibroblast growth factor 4 and oncostatin M. In vivo, MSC were transplanted into spleen or liver of NOD/SCID mice undergoing partial hepatectomy and retrorsine treatment. Expression of mesenchymal and hepatic markers was analyzed by RT-PCR, Western blot and immunohistochemistry. In vitro, adult and pediatric MSC expressed characteristic surface antigens of MSC. Expansion capacity of pediatric MSC was significantly higher when compared to adult MSC. In co-culture with Huh-7 cells in hepatogenic differentiation medium, albumin expression was more frequently detected in pediatric MSC (5/8 experiments) when compared to adult MSC (2/10 experiments). However, in such condition pediatric MSC expressed alpha smooth muscle more strongly than adult MSC. Stable engraftment in the liver was not achieved after intrasplenic injection of pediatric or adult MSC. After intrahepatic injection, MSC permanently remained in liver tissue, kept a mesenchymal morphology and expressed vimentin and alpha smooth muscle actin, but no hepatic markers. Further, MSC localization merges with collagen deposition in transplanted liver and no difference was observed using adult or pediatric MSC. In conclusion, when transplanted into an injured or regenerating liver, MSC differentiated into myofibroblasts with development of fibrous tissue, regardless of donor age. These results indicate that MSC in certain circumstances might be harmful due to their fibrogenic potential and this should be considered before potential use of MSC for cell therapy.

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Engrafted MSC express alpha smooth muscle actin and merge with collagen deposition in mouse liver.Eight weeks after injection, sections from MSC injected (A) or sham injected (B) livers were stained with an Ab specific against human vimentin (panels a) or an Ab against αSMA (panels b) or with Hoechst (panels e). MSC retained their spindle like morphology and formed band-like structures similar to fibrotic livers. Overlay images of Hoechst and vimentin (panels c) or Hoechst and αSMA (panels d) are shown. Serial sections were stained with Masson to visualize collagen deposition (blue staining). Comparing collagen stained by Masson (blue) with staining of vimentin and αSMA demonstrate similar localization (Aa, Ab, Af).
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pone-0006657-g007: Engrafted MSC express alpha smooth muscle actin and merge with collagen deposition in mouse liver.Eight weeks after injection, sections from MSC injected (A) or sham injected (B) livers were stained with an Ab specific against human vimentin (panels a) or an Ab against αSMA (panels b) or with Hoechst (panels e). MSC retained their spindle like morphology and formed band-like structures similar to fibrotic livers. Overlay images of Hoechst and vimentin (panels c) or Hoechst and αSMA (panels d) are shown. Serial sections were stained with Masson to visualize collagen deposition (blue staining). Comparing collagen stained by Masson (blue) with staining of vimentin and αSMA demonstrate similar localization (Aa, Ab, Af).

Mentions: During fibrosis, myofibroblasts expressing αSMA appear within the liver. Recently, it has been shown that these cells can be of bone marrow origin [25]. Therefore we investigated whether transplanted MSC could differentiate in myofibroblasts. Staining for αSMA on sections of transplanted mouse liver showed that MSC express αSMA (Fig. 7A). Antibody against αSMA is not human specific and stains smooth muscle cells around blood vessels of mouse livers as shown on liver sections of sham injected mice (Fig. 7B, b and d). Histochemistry using Masson's trichrome on serial sections showed that collagen deposition merges with vimentin- and αSMA-expressing MSC (Fig. 7A, a, b and f).


Fibrogenic potential of human multipotent mesenchymal stromal cells in injured liver.

Baertschiger RM, Serre-Beinier V, Morel P, Bosco D, Peyrou M, Clément S, Sgroi A, Kaelin A, Buhler LH, Gonelle-Gispert C - PLoS ONE (2009)

Engrafted MSC express alpha smooth muscle actin and merge with collagen deposition in mouse liver.Eight weeks after injection, sections from MSC injected (A) or sham injected (B) livers were stained with an Ab specific against human vimentin (panels a) or an Ab against αSMA (panels b) or with Hoechst (panels e). MSC retained their spindle like morphology and formed band-like structures similar to fibrotic livers. Overlay images of Hoechst and vimentin (panels c) or Hoechst and αSMA (panels d) are shown. Serial sections were stained with Masson to visualize collagen deposition (blue staining). Comparing collagen stained by Masson (blue) with staining of vimentin and αSMA demonstrate similar localization (Aa, Ab, Af).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2722022&req=5

pone-0006657-g007: Engrafted MSC express alpha smooth muscle actin and merge with collagen deposition in mouse liver.Eight weeks after injection, sections from MSC injected (A) or sham injected (B) livers were stained with an Ab specific against human vimentin (panels a) or an Ab against αSMA (panels b) or with Hoechst (panels e). MSC retained their spindle like morphology and formed band-like structures similar to fibrotic livers. Overlay images of Hoechst and vimentin (panels c) or Hoechst and αSMA (panels d) are shown. Serial sections were stained with Masson to visualize collagen deposition (blue staining). Comparing collagen stained by Masson (blue) with staining of vimentin and αSMA demonstrate similar localization (Aa, Ab, Af).
Mentions: During fibrosis, myofibroblasts expressing αSMA appear within the liver. Recently, it has been shown that these cells can be of bone marrow origin [25]. Therefore we investigated whether transplanted MSC could differentiate in myofibroblasts. Staining for αSMA on sections of transplanted mouse liver showed that MSC express αSMA (Fig. 7A). Antibody against αSMA is not human specific and stains smooth muscle cells around blood vessels of mouse livers as shown on liver sections of sham injected mice (Fig. 7B, b and d). Histochemistry using Masson's trichrome on serial sections showed that collagen deposition merges with vimentin- and αSMA-expressing MSC (Fig. 7A, a, b and f).

Bottom Line: Expansion capacity of pediatric MSC was significantly higher when compared to adult MSC.In conclusion, when transplanted into an injured or regenerating liver, MSC differentiated into myofibroblasts with development of fibrous tissue, regardless of donor age.These results indicate that MSC in certain circumstances might be harmful due to their fibrogenic potential and this should be considered before potential use of MSC for cell therapy.

View Article: PubMed Central - PubMed

Affiliation: Surgical Research Unit, Department of Surgery, University Hospital Geneva, Geneva, Switzerland.

ABSTRACT
Multipotent mesenchymal stromal cells (MSC) are currently investigated clinically as cellular therapy for a variety of diseases. Differentiation of MSC toward endodermal lineages, including hepatocytes and their therapeutic effect on fibrosis has been described but remains controversial. Recent evidence attributed a fibrotic potential to MSC. As differentiation potential might be dependent of donor age, we studied MSC derived from adult and pediatric human bone marrow and their potential to differentiate into hepatocytes or myofibroblasts in vitro and in vivo. Following characterization, expanded adult and pediatric MSC were co-cultured with a human hepatoma cell line, Huh-7, in a hepatogenic differentiation medium containing Hepatocyte growth factor, Fibroblast growth factor 4 and oncostatin M. In vivo, MSC were transplanted into spleen or liver of NOD/SCID mice undergoing partial hepatectomy and retrorsine treatment. Expression of mesenchymal and hepatic markers was analyzed by RT-PCR, Western blot and immunohistochemistry. In vitro, adult and pediatric MSC expressed characteristic surface antigens of MSC. Expansion capacity of pediatric MSC was significantly higher when compared to adult MSC. In co-culture with Huh-7 cells in hepatogenic differentiation medium, albumin expression was more frequently detected in pediatric MSC (5/8 experiments) when compared to adult MSC (2/10 experiments). However, in such condition pediatric MSC expressed alpha smooth muscle more strongly than adult MSC. Stable engraftment in the liver was not achieved after intrasplenic injection of pediatric or adult MSC. After intrahepatic injection, MSC permanently remained in liver tissue, kept a mesenchymal morphology and expressed vimentin and alpha smooth muscle actin, but no hepatic markers. Further, MSC localization merges with collagen deposition in transplanted liver and no difference was observed using adult or pediatric MSC. In conclusion, when transplanted into an injured or regenerating liver, MSC differentiated into myofibroblasts with development of fibrous tissue, regardless of donor age. These results indicate that MSC in certain circumstances might be harmful due to their fibrogenic potential and this should be considered before potential use of MSC for cell therapy.

Show MeSH
Related in: MedlinePlus