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Glycobiology Advance Access originally published online on May 28, 2003
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Glycobiology, 2003, Vol. 13, No. 9 647-653
© 2003 Oxford University Press

Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies

Isabelle Barbosa2, Stéphanie Garcia2, Véronique Barbier-Chassefière3, Jean-Pierre Caruelle2, Isabelle Martelly2 and Dulce Papy-García1,2

2 Laboratoire de Recherche sur la Croissance, la Réparation et la Régénération Tissulaires (CRRET), CNRS FRE-2412, Université Paris 12-Val de Marne, 61 Avenue du Général de Gaulle, 94010 Créteil cedex, France
3 Société OTR3 SARL, 33, avenue Pierre Brossolette, 94000 Créteil cedex, France

Received on February 21, 2003; revised on April 14, 2003; accepted on May 8, 2003

This article describes a simple and selective procedure used for direct measurement of sulfated glycosaminoglycans (GAGs) in biological samples and its application to the determination of GAGs during tissue regeneration and myogenic differentiation. We describe a modified procedure of previous GAG assays that has improved specificity, reproducibility, and sensitivity. The assay is based on the ability of sulfated GAGs to bind the cationic dye 1,9-dimethylmethylene blue. We describe conditions that allow isolation of the GAG–dye complex. This complex was dissociated; the optical density measurement of the dissociated dye permitted quantification of GAGs in biological samples. Applied to the study of myogenic cell differentiation in vitro, muscle repair, and skin ulceration, this method revealed significant modifications in the patterns of expression of different sulfated GAGs in these tissues. In particular, application of the method after nitrous acid treatment revealed that heparan sulfate and chondroitin sulfate ratio changed during muscle regeneration process.

1 To whom correspondence should be addressed; e-mail: papy{at}univ-paris12.fr


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