Analysis of Proteoglycan Synthesis and Secretion in Cell Culture Systems

Chiara Paganini; Rossella Costantini; Antonio Rossi

doi:10.1007/978-1-4939-9133-4_6

Analysis of Proteoglycan Synthesis and Secretion in Cell Culture Systems

Methods Mol Biol. 2019:1952:71-80. doi: 10.1007/978-1-4939-9133-4_6.

Authors

Chiara Paganini¹, Rossella Costantini¹, Antonio Rossi²

Affiliations

¹ Department of Molecular Medicine, Biochemistry Unit, University of Pavia, Pavia, Italy.
² Department of Molecular Medicine, Biochemistry Unit, University of Pavia, Pavia, Italy. antrossi@unipv.it.

PMID: 30825166
DOI: 10.1007/978-1-4939-9133-4_6

Abstract

Experimental protocols for the synthesis and secretion of proteoglycans in cell culture models are important to study specific biosynthetic steps or disorders in which a defect in proteoglycans is expected. We describe a method using ³⁵S-sulfate to metabolically label newly synthesized proteoglycans from cell cultures in order to measure proteoglycan synthesis and secretion. The method is set up for fibroblast and chondrocyte cultures, but can be extended to other cell types.

Keywords: Gel filtration chromatography; Glycosaminoglycan; Ion-exchange chromatography; Metabolic labeling; Protein secretion; Proteoglycan.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biosynthetic Pathways
Cell Culture Techniques / methods*
Chondrocytes / chemistry
Chondrocytes / metabolism
Chromatography, Gel / methods*
Chromatography, Ion Exchange / methods*
Fibroblasts / chemistry
Fibroblasts / metabolism
Humans
Proteoglycans / analysis*
Proteoglycans / metabolism

Substances

Proteoglycans