The Formation of Extracellular Matrix During Chondrogenic Differentiation of Mesenchymal Stem Cells Correlates with Increased Levels of Xylosyltransferase I (original) (raw)

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,

Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

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Institut für Transfusionsmedizin und Immunologie, DRK‐Blutspendedienst Baden‐Württemberg‐Hessen

, Mannheim,

Germany

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Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

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,

Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

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,

Institut für Transfusionsmedizin und Immunologie, DRK‐Blutspendedienst Baden‐Württemberg‐Hessen

, Mannheim,

Germany

Search for other works by this author on:

,

Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

Search for other works by this author on:

,

Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

Search for other works by this author on:

,

Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

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Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum NRW, Universitätsklinik der Ruhr‐Universität Bochum

, Bad Oeynhausen,

Germany

Correspondence: Christian Götting, Ph.D., Institut für Laboratoriums‐ und Transfusionsmedizin, Herz‐ und Diabeteszentrum NRW, Georgstraβe 11, 32545 Bad Oeynhausen, Germany. Telephone: +49‐5731‐972033; Fax: +49‐5731‐972013; e-mail: cgoetting@hdz‐nrw.de

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Received:

12 October 2005

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Christian Prante, Karen Bieback, Carsten Funke, Sylvia Schön, Susanne Kern, Joachim Kuhn, Martin Gastens, Knut Kleesiek, Christian Götting, The Formation of Extracellular Matrix During Chondrogenic Differentiation of Mesenchymal Stem Cells Correlates with Increased Levels of Xylosyltransferase I, Stem Cells, Volume 24, Issue 10, October 2006, Pages 2252–2261, https://doi.org/10.1634/stemcells.2005-0508
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Abstract

In vitro differentiation of mesenchymal stem cells (MSCs) into chondrogenic cells and their transplantation is promising as a technique for the treatment of cartilaginous defects. But the regulation of extracellular matrix (ECM) formation remains elusive. Therefore, the objective of this study was to analyze the regulation of proteoglycan (PG) biosynthesis during the chondrogenic differentiation of MSCs. In different stages of chondrogenic differentiation, we analyzed mRNA and protein expression of key enzymes and PG core proteins involved in ECM development. For xylosyltransferase I (XT‐I), we found maximum mRNA levels 48 hours after chondrogenic induction with a 5.04 ± 0.58 (mean ± SD)‐fold increase. This result correlates with significantly elevated levels of enzymatic XT‐I activity (0.49 ± 0.03 μU/1 × 106 cells) at this time point. Immunohistochemical staining of XT‐I revealed a predominant upregulation in early chondrogenic stages. The highly homologous protein XT‐II showed 4.7‐fold (SD 0.6) increased mRNA levels on day 7. To determine the differential expression of heparan sulfate (HS), chondroitin sulfate (CS), and dermatan sulfate (DS) chains, we analyzed the mRNA expression of EXTL2 (α‐4‐_N_‐acetylhexosaminyltransferase), GalNAcT (β‐1,4‐_N_‐acetylgalactosaminyltransferase), and GlcAC5E (glucuronyl C5 epimerase). All key enzymes showed a similar regulation with temporarily downregulated mRNA levels (up to −87‐fold) after chondrogenic induction. In accordance to previous studies, we observed a similar increase in the expression of PG core proteins. In conclusion, we could show that key enzymes for CS, DS, and HS synthesis, especially XT‐I, are useful markers for the developmental stages of chondrogenic differentiation.

Copyright © 2006 AlphaMed Press

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