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Isolation of Astrocytes Displaying Myofibroblast Properties and Present in Multiple Sclerosis Lesions

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Abstract

A wide heterogeneity of lesions can affect the central nervous system (CNS). In all situations where neurons are damaged, including multiple sclerosis (MS), a common reactive astrocytosis is present. Sedimentation field-flow fractionation (SdFFF) was used to sort astrocyte subpopulations. After SdFFF elution, cells, prepared from rat newborn cortex, were cultured and analyzed by immunocytofluorescence for glial fibrillary acidic protein (GFAP) and α-smooth muscle (SM) actin (a specific marker for myofibroblasts) expression. Cell contractile capacity was studied. Samples from patients with MS were also analyzed. Three main fractions (F1, F2, and F3) were isolated and compared with the total eluted population (TP). TP, F1, F2, and F3, contained respectively 74, 96, 12, and 98% of GFAP expressing astrocytes. In F3, astrocytes only expressed GFAP while in F1, astrocytes expressed both GFAP and α-SM actin. In F2 and TP, α-SM actin expression was barely detected. F3-derived cells showed higher contractile capacities compared with F1-derived cells. In one specific case of MS known as Baló’s concentric MS, astrocytes expressing both GFAP and α-SM actin were detected. Using SdFFF, a population of astrocytes presenting myofibroblast properties was isolated. This subpopulation of astrocytes was also observed in a MS sample suggesting that it could be involved in lesion formation and remodeling during CNS pathologies.

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Acknowledgements

The authors kindly thank Claire Carrion (University of Limoges, Confocal microscopy facility, Limoges, France) for confocal microscopy analysis. The authors also thank the Department of Neurology of the University Hospital of Limoges (France) for providing human samples. Nicolas Vedrenne and Vincent Sarrazy were supported by fellowships from the “Conseil Régional du Limousin”.

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Author notes

  1. Vincent Sarrazy

    Present address: Mediterranean Centre for Molecular Medicine, University of Nice Sophia Antipolis, INSERM U1065, 06000, Nice, France

Authors and Affiliations

  1. EA 6309 “Myeline maintenance and peripheral neuropathies”, University of Limoges, 87000, Limoges, France

    Nicolas Vedrenne, Vincent Sarrazy, Laurence Richard, Nelly Bordeau, Fabrice Billet & Alexis Desmoulière

  2. Department of Neurology, University Hospital of Limoges, 87000, Limoges, France

    Laurence Richard

  3. EA 3842 “Cellular homeostasis and pathologies”, University of Limoges, 87000, Limoges, France

    Serge Battu

  4. Department of Physiology, Faculty of Pharmacy, University of Limoges, 2 rue du Dr. Marcland, 87025, Limoges Cedex, France

    Alexis Desmoulière

Authors
  1. Nicolas Vedrenne
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  2. Vincent Sarrazy
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  4. Nelly Bordeau
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  5. Serge Battu
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  7. Alexis Desmoulière
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Correspondence to Alexis Desmoulière.

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Vedrenne, N., Sarrazy, V., Richard, L. et al. Isolation of Astrocytes Displaying Myofibroblast Properties and Present in Multiple Sclerosis Lesions. Neurochem Res 42, 2427–2434 (2017). https://doi.org/10.1007/s11064-017-2268-y

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  • DOI: https://doi.org/10.1007/s11064-017-2268-y

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