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Anti‐ACSA‐2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes
Authors:Christina G. Kantzer  Camille Boutin  Ina D. Herzig  Carolina Wittwer  Sandy Reiß  Marie Catherine Tiveron  Jan Drewes  Thomas D. Rockel  Stefanie Ohlig  Jovica Ninkovic  Harold Cremer  Sandra Pennartz  Melanie Jungblut  Andreas Bosio
Affiliation:1. Miltenyi Biotec GmbH, Friedrich‐Ebert‐Straße 68, Bergisch Gladbach, 51429 Germany;2. Aix‐Marseille Université, CNRS, Institut de Biologie du Développement de Marseille, Campus de Luminy, Marseille, 13288 France;3. Institute of Stem Cell Research, Helmholtz Center Munich, German Research Center for Environmental Health, Großhadernerstr.9, Planegg/Munich, 82152 Germany;4. Physiological Genomics, Biomedical Center, Ludwig‐Maximilian University Munich, Großhadernerstr.9, Planegg/Munich, 82152 Germany;5. Miltenyi Biotec GmbH, Friedrich‐Ebert‐Straße 68, Bergisch Gladbach, 51429 GermanyCorrespondence Andreas Bosio, Miltenyi Biotec GmbH, 51429 Bergisch Gladbach, Germany. Email:
Abstract:Astrocytes are the most abundant cell type of the central nervous system and cover a broad range of functionalities. We report here the generation of a novel monoclonal antibody, anti‐astrocyte cell surface antigen‐2 (Anti‐ACSA‐2). Flow cytometry, immunohistochemistry and immunocytochemistry revealed that Anti‐ACSA‐2 reacted specifically with a not yet identified glycosylated surface molecule of murine astrocytes at all developmental stages. It did not show any labeling of non‐astroglial cells such as neurons, oligodendrocytes, NG2+ cells, microglia, endothelial cells, leukocytes, or erythrocytes. Co‐labeling studies of GLAST and ACSA‐2 showed largely overlapping expression. However, there were also notable differences in protein expression levels and frequencies of single‐positive subpopulations of cells in some regions of the CNS such as cerebellum, most prominently at early postnatal stages. In the neurogenic niches, the dentate gyrus of the hippocampus and the subventricular zone (SVZ), again a general overlap with slight differences in expression levels were observed. ACSA‐2 was unlike GLAST not sensitive to papain‐based tissue dissociation and allowed for a highly effective, acute, specific, and prospective purification of viable astrocytes based on a new rapid sorting procedure using Anti‐ACSA‐2 directly coupled to superparamagnetic MicroBeads. In conclusion, ACSA‐2 appears to be a new surface marker for astrocytes, radial glia, neural stem cells and bipotent glial progenitor cells which opens up the possibility of further dissecting the characteristics of astroglial subpopulations and lineages.
Keywords:astrocyte cell surface antigen‐2  antibody  cell separation  cell surface marker  GLAST
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