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Glycosaminoglycan profiling in different cell types using infrared spectroscopy and imaging

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Abstract

We recently identified vibrational spectroscopic markers characteristic of standard glycosaminoglycan (GAG) molecules. The aims of the present work were to further this investigation to more complex biological systems and to characterize, via their spectral profiles, cell types with different capacities for GAG synthesis. After recording spectral information from individual GAG standards (hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate) and GAG-GAG mixtures, GAG-defective mutant Chinese hamster ovary (CHO)-745 cells, wild-type CHO cells, and chondrocytes were analyzed as suspensions by high-throughput infrared spectroscopy and as single isolated cells by infrared imaging. Spectral data were processed and interpreted by exploratory unsupervised chemometric methods based on hierarchical cluster analysis and principal component analysis. Our results showed that the spectral information obtained was discriminant enough to clearly delineate between the different cell types both at the cell suspension and single-cell levels. The abilities of the technique are to perform spectral profiling and to identify single cells with different potentials to synthesize GAGs. Infrared microspectroscopy/imaging could therefore be developed for cell screening purposes and further for identifying GAG molecules in normal tissues during physiological conditions (aging, healing process) and numerous pathological states (arthritis, cancer).

FTIR imaging for profiling GAG-synthesizing cells

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Abbreviations

CHO:

Chinese hamster ovary

CS:

Chondroitin sulfate

DPBS:

Dulbecco’s phosphate buffer saline

DS:

Dermatan sulfate

FTIR:

Fourier transform infrared

GAG:

Glycosaminoglycan

HA:

Hyaluronic acid

HCA:

Hierarchical cluster analysis

HEP:

Heparin

HS:

Heparan sulfate

KS:

Keratan sulfate

PC:

Principal component

PCA:

Principal component analysis

PFA:

Paraformaldehyde

PG:

Proteoglycan

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Acknowledgments

The authors thank the Reims PICT-IBiSA Platform and the Service de Chirurgie Orthopédique, CHU de Reims. The financial support of the Ligue Nationale contre le Cancer (Comité de la Marne et Comité de Haute-Marne, Conférence de Coordination InterRégionale du Grand Est (CCIR-GE)), the FEDER and the Région Champagne-Ardenne (CPER 2007–2013), FIS (contract PI10/15), and RTICC (Red Temática de Investigación Cooperativa en Cáncer, contracts RD06/0020/0102; RD12/0036/0020), Instituto Carlos III Madrid & ERDF (European Regional Development Fund), Spain, is gratefully acknowledged.

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Authors and Affiliations

  1. Laboratoire de Biochimie médicale et de Biologie Moléculaire, UFR de Médecine, Université de Reims Champagne-Ardenne, 51 rue Cognacq-Jay, 51095, Reims Cedex, France

    Stéphane Brézillon, Lila Lovergne, Irene Tadeo, François-Xavier Maquart & Yanusz Wegrowski

  2. CNRS UMR7369, Matrice Extracellulaire et Dynamique Cellulaire, MEDyC, 51095, Reims, France

    Stéphane Brézillon, Valérie Untereiner, François-Xavier Maquart, Yanusz Wegrowski & Ganesh D. Sockalingum

  3. MéDIAN-Biophotonique et Technologies pour la Santé, UFR de Pharmacie, Université de Reims Champagne-Ardenne, 51 rue Cognacq-Jay, 51096, Reims Cedex, France

    Valérie Untereiner, Lila Lovergne & Ganesh D. Sockalingum

  4. Laboratoire Central de Biochimie, CHU de Reims, 51092, Reims Cedex, France

    François-Xavier Maquart

  5. Laboratorio de Patología Molecular, Departamento de Patología, Facultad de Medicina y Odontología, INCLIVA (Universidad de Valencia; Fundación Investigatión Hospital Clínico de Valencia), Avda. Blasco Ibañez 15, Valencia, 46010, Spain

    Irene Tadeo & Rosa Noguera

Authors
  1. Stéphane Brézillon
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  2. Valérie Untereiner
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  3. Lila Lovergne
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  4. Irene Tadeo
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  5. Rosa Noguera
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  6. François-Xavier Maquart
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  7. Yanusz Wegrowski
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  8. Ganesh D. Sockalingum
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Corresponding author

Correspondence to Ganesh D. Sockalingum.

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Brézillon, S., Untereiner, V., Lovergne, L. et al. Glycosaminoglycan profiling in different cell types using infrared spectroscopy and imaging. Anal Bioanal Chem 406, 5795–5803 (2014). https://doi.org/10.1007/s00216-014-7994-2

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  • DOI: https://doi.org/10.1007/s00216-014-7994-2

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