Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes.
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.
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Christina G
CG
Kantzer
Aix-Marseille Université, CNRS, Institut de Biologie du Développement de Marseille, Campus de Luminy, Marseille, 13288, France.
Camille
C
Boutin
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Ina D
ID
Herzig
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Carolina
C
Wittwer
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Sandy
S
Reiß
Aix-Marseille Université, CNRS, Institut de Biologie du Développement de Marseille, Campus de Luminy, Marseille, 13288, France.
Marie Catherine
MC
Tiveron
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Jan
J
Drewes
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Thomas D
TD
Rockel
Institute of Stem Cell Research, Helmholtz Center Munich, German Research Center for Environmental Health, Großhadernerstr.9, Planegg/Munich, 82152, Germany.|Physiological Genomics, Biomedical Center, Ludwig-Maximilian University Munich, Großhadernerstr.9, Planegg/Munich, 82152, Germany.
Stefanie
S
Ohlig
Institute of Stem Cell Research, Helmholtz Center Munich, German Research Center for Environmental Health, Großhadernerstr.9, Planegg/Munich, 82152, Germany.|Physiological Genomics, Biomedical Center, Ludwig-Maximilian University Munich, Großhadernerstr.9, Planegg/Munich, 82152, Germany.
Jovica
J
Ninkovic
Aix-Marseille Université, CNRS, Institut de Biologie du Développement de Marseille, Campus de Luminy, Marseille, 13288, France.
Harold
H
Cremer
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Sandra
S
Pennartz
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Melanie
M
Jungblut
Miltenyi Biotec GmbH, Friedrich-Ebert-Straße 68, Bergisch Gladbach, 51429, Germany.
Andreas
0000-0001-9776-2535
A
Bosio
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D001253:Astrocytes / Q000166:cytology* / Q000276:immunology*; D001921:Brain / Q000166:cytology / Q000254:growth & development; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D027341:Excitatory Amino Acid Transporter 1 / Q000032:analysis; D018189:Immunomagnetic Separation / Q000379:methods*; D007962:Leukocytes / Q000166:cytology / Q000276:immunology; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D017628:Microglia / Q000166:cytology / Q000276:immunology; D058953:Neural Stem Cells / Q000276:immunology; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000276:immunology; D017208:Rats, Wistar
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D027341:Excitatory Amino Acid Transporter 1; C493393:Slc1a3 protein, mouse
GLAST; antibody; astrocyte cell surface antigen-2; cell separation; cell surface marker