title: Probing glycosaminoglycan spectral signatures in live cells and their conditioned media by Raman microspectroscopy.
abstract: 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.
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, Laboratoire de Biochimie médicale et de Biologie Moléculaire, UFR de Médecine, Reims, France.
authors.forename: S
authors.initials: S
authors.lastname: Brézillon
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, MéDIAN-Biophotonique et Technologies pour la Santé, UFR de Pharmacie, Reims, France and Université de Reims Champagne-Ardenne, Plateforme d'imagerie cellulaire et tissulaire (PICT), Reims, France.
authors.forename: V
authors.initials: V
authors.lastname: Untereiner
authors.affiliation: Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt.
authors.forename: H T
authors.initials: HT
authors.lastname: Mohamed
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, Laboratoire de Biochimie médicale et de Biologie Moléculaire, UFR de Médecine, Reims, France and Université de Reims Champagne-Ardenne, MéDIAN-Biophotonique et Technologies pour la Santé, UFR de Pharmacie, Reims, France.
authors.forename: J
authors.initials: J
authors.lastname: Hodin
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, Laboratoire de Biochimie médicale et de Biologie Moléculaire, UFR de Médecine, Reims, France.
authors.forename: A
authors.initials: A
authors.lastname: Chatron-Colliet
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, Laboratoire de Biochimie médicale et de Biologie Moléculaire, UFR de Médecine, Reims, France and Laboratoire Central de Biochimie, CHU de Reims, Reims, France.
authors.forename: F-X
authors.initials: FX
authors.lastname: Maquart
authors.affiliation: CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire (MEDyC), Reims, France. stephane.brezillon@univ-reims.fr ganesh.sockalingum@univ-reims.fr and Université de Reims Champagne-Ardenne, MéDIAN-Biophotonique et Technologies pour la Santé, UFR de Pharmacie, Reims, France.
authors.forename: G D
authors.identifier: http://orcid.org/0000-0001-7954-8340
authors.initials: GD
authors.lastname: Sockalingum
mesh_terms: D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D019902:Chondrocytes / Q000166:cytology*; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology*; D006025:Glycosaminoglycans / Q000737:chemistry*; D006801:Humans; D008545:Melanoma; D013059:Spectrum Analysis, Raman
chemical_list: D017077:Culture Media, Conditioned; D006025:Glycosaminoglycans