Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs

Chondrogenic differentiating mesenchymal stem cells (MSCs) are mimicking embryonal endochondral ossification and become hypertrophic. BMP (bone morphogenetic protein) and Activin Membrane Bound Inhibitor (BAMBI) is a pseudoreceptor that regulates the activity of transforming growth factor-β (TGF-β)...

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Main Authors: Christian G. Pfeifer, Alexandra Karl, Arne Berner, Johannes Zellner, Paul Schmitz, Markus Loibl, Matthias Koch, Peter Angele, Michael Nerlich, Michael B. Mueller
Format: Article
Language:English
Published: Wiley 2016-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2016/2685147
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author Christian G. Pfeifer
Alexandra Karl
Arne Berner
Johannes Zellner
Paul Schmitz
Markus Loibl
Matthias Koch
Peter Angele
Michael Nerlich
Michael B. Mueller
author_facet Christian G. Pfeifer
Alexandra Karl
Arne Berner
Johannes Zellner
Paul Schmitz
Markus Loibl
Matthias Koch
Peter Angele
Michael Nerlich
Michael B. Mueller
author_sort Christian G. Pfeifer
collection DOAJ
description Chondrogenic differentiating mesenchymal stem cells (MSCs) are mimicking embryonal endochondral ossification and become hypertrophic. BMP (bone morphogenetic protein) and Activin Membrane Bound Inhibitor (BAMBI) is a pseudoreceptor that regulates the activity of transforming growth factor-β (TGF-β) and BMP signalling during chondrogenesis. Both TGF-β and BMP signalling are regulators of chondrogenic cell differentiation. Human bone marrow derived MSCs were chondrogenically predifferentiated in aggregate culture for 14 days. Thereafter, one group was subjected to hypertrophy enhancing media conditions while controls were kept in chondrogenic medium until day 28. Histological evaluation, gene expression by PCR, and Western blot analysis were carried out at days 1, 3, 7, 14, 17, 21, and 28. A subset of cultures was treated with the BMP inhibitor Noggin to test for BMP dependent expression of BAMBI. Hypertrophic differentiated pellets showed larger cells with increased collagen 10 and alkaline phosphatase staining. There was significantly increased expression of BAMBI on gene expression and protein level in hypertrophic cultures compared to the chondrogenic control and increased BMP4 gene expression. Immunohistochemistry showed intense staining of BAMBI in hypertrophic cells. BAMBI expression was dose-dependently downregulated by Noggin. The pseudoreceptor BAMBI is upregulated upon enhancement of hypertrophy in MSC chondrogenic differentiation by a BMP dependent mechanism.
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spelling doaj-art-e2644eaa3e004610b86d2c661efe664e2025-08-20T02:21:14ZengWileyStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/26851472685147Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCsChristian G. Pfeifer0Alexandra Karl1Arne Berner2Johannes Zellner3Paul Schmitz4Markus Loibl5Matthias Koch6Peter Angele7Michael Nerlich8Michael B. Mueller9Department of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyDepartment of Trauma and Orthopaedic Surgery, University Clinic Regensburg, Regensburg, GermanyChondrogenic differentiating mesenchymal stem cells (MSCs) are mimicking embryonal endochondral ossification and become hypertrophic. BMP (bone morphogenetic protein) and Activin Membrane Bound Inhibitor (BAMBI) is a pseudoreceptor that regulates the activity of transforming growth factor-β (TGF-β) and BMP signalling during chondrogenesis. Both TGF-β and BMP signalling are regulators of chondrogenic cell differentiation. Human bone marrow derived MSCs were chondrogenically predifferentiated in aggregate culture for 14 days. Thereafter, one group was subjected to hypertrophy enhancing media conditions while controls were kept in chondrogenic medium until day 28. Histological evaluation, gene expression by PCR, and Western blot analysis were carried out at days 1, 3, 7, 14, 17, 21, and 28. A subset of cultures was treated with the BMP inhibitor Noggin to test for BMP dependent expression of BAMBI. Hypertrophic differentiated pellets showed larger cells with increased collagen 10 and alkaline phosphatase staining. There was significantly increased expression of BAMBI on gene expression and protein level in hypertrophic cultures compared to the chondrogenic control and increased BMP4 gene expression. Immunohistochemistry showed intense staining of BAMBI in hypertrophic cells. BAMBI expression was dose-dependently downregulated by Noggin. The pseudoreceptor BAMBI is upregulated upon enhancement of hypertrophy in MSC chondrogenic differentiation by a BMP dependent mechanism.http://dx.doi.org/10.1155/2016/2685147
spellingShingle Christian G. Pfeifer
Alexandra Karl
Arne Berner
Johannes Zellner
Paul Schmitz
Markus Loibl
Matthias Koch
Peter Angele
Michael Nerlich
Michael B. Mueller
Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
Stem Cells International
title Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
title_full Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
title_fullStr Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
title_full_unstemmed Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
title_short Expression of BMP and Actin Membrane Bound Inhibitor Is Increased during Terminal Differentiation of MSCs
title_sort expression of bmp and actin membrane bound inhibitor is increased during terminal differentiation of mscs
url http://dx.doi.org/10.1155/2016/2685147
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