Probing glycosaminoglycan spectral signatures in live cells and their conditioned media by Raman microspectroscopy.
Fiche publication
Date publication
mars 2017
Journal
The Analyst
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BREZILLON Stéphane, Pr SOCKALINGUM Ganesh
Tous les auteurs :
Brézillon S, Untereiner V, Mohamed HT, Hodin J, Chatron-Colliet A, Maquart FX, Sockalingum GD
Lien Pubmed
Résumé
Spectroscopic markers characteristic of reference glycosaminoglycan molecules were identified previously based on their vibrational signatures. Infrared spectral signatures of glycosaminoglycans in fixed cells were also recently demonstrated but probing live cells still remains challenging. Raman microspectroscopy is potentially interesting to perform studies under physiological conditions. The aim of the present work was to identify the Raman spectral signatures of GAGs in fixed and live cells and in their conditioned media. Biochemical and Raman analyses were performed on five cell types: chondrocytes, dermal fibroblasts, melanoma (SK-MEL-28), wild type CHO, and glycosaminoglycan-defective mutant CHO-745 cells. The biochemical assay of sulfated GAGs in conditioned media was only possible for chondrocytes, dermal fibroblasts, and wild type CHO due to the detection limit of the test. In contrast, Raman microspectroscopy allowed probing total glycosaminoglycan content in conditioned media, fixed and live cells and the data were analysed by principal component analysis. Our results showed that the Raman technique is sensitive enough to identify spectral markers of glycosaminoglycans that were useful to characterise the conditioned media of the five cell types. The results were confirmed at the single cell level on both live and fixed cells with a good differentiation between the cell types. Furthermore, the principal component loadings revealed prominent glycosaminoglycan-related spectral information. Raman microspectroscopy allows monitoring of the glycosaminoglycan profiles of single live cells and could therefore be developed for cell screening purposes and holds promise for identifying glycosaminoglycan signatures as a marker of cancer progression in tissues.
Mots clés
Animals, CHO Cells, Cell Line, Tumor, Chondrocytes, cytology, Cricetinae, Cricetulus, Culture Media, Conditioned, chemistry, Fibroblasts, cytology, Glycosaminoglycans, chemistry, Humans, Melanoma, Spectrum Analysis, Raman
Référence
Analyst. 2017 Mar;: