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First published online June 15, 2006
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Submitted on October 12, 2005
Accepted on June 10, 2006

Tissue-Specific Stem Cells

The formation of extracellular matrix during chondrogenic differentiation of mesenchymal stem cells correlates with increased levels of xylosyltransferase I

Christian Prante 1, Karen Bieback 2, Carsten Funke 1, Sylvia Schön 1, Susanne Kern 2, Joachim Kuhn 1, Martin Gastens 1, Knut Kleesiek 1, Christian Götting 1*

1 Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany
2 Institut für Transfusionsmedizin und Immunologie, DRK-Blutspendedienst Baden-Württemberg - Hessen, Mannheim, Germany

* To whom correspondence should be addressed. E-mail: cgoetting{at}hdz-nrw.de.


  Abstract

In vitro differentiation of mesenchymal stem cells (MSCs) into chondrogenic cells and their transplantation are a promising technique for the treatment of cartilaginous defects. But the regulation of extracellular matrix (ECM) formation remains still elusive. Therefore, the objective of this study was to analyze the regulation of proteoglycan biosynthesis during the chondrogenic differentiation of MSCs. In different stages of chondrogenic differentiation we analyzed mRNA and protein expression of key enzymes and proteoglycan core proteins involved in the ECM development.

For xylosyltransferase I (XT-I) we found maximum mRNA levels 48 h 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/1x106 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 (HS), chondroitin (CS) and dermatan (DS) sulfate chains we analyzed the mRNA expression of {alpha}4-Nacetylhexosaminyltransferase (EXTL2), beta1,4-N-acetylgalactosaminyltransferase (GalNAcT) and glucuronyl C5 epimerase (GlcAC5E). All key enzymes showed a similar regulation with temporarily down-regulated mRNA levels (up to -87-fold) after chondrogenic induction. In accordance to previous studies we observed a similar increase in the expression of proteoglycan 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.


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 EndocrinologyHome page
A. Woods, S. Khan, and F. Beier
C-Type Natriuretic Peptide Regulates Cellular Condensation and Glycosaminoglycan Synthesis during Chondrogenesis
Endocrinology, October 1, 2007; 148(10): 5030 - 5041.
[Abstract] [Full Text] [PDF]


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