{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T15:28:09Z","timestamp":1787498889334,"version":"build-2736575974"},"reference-count":29,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,2,18]],"date-time":"2026-02-18T00:00:00Z","timestamp":1771372800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Astrocyte-specific cell surface antigen-2 (ACSA2) has been established as the gold-standard marker for isolating astrocytes via magnetic-activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS) for downstream transcriptomic studies. In a prior study of the astrocyte response to cortical stroke, we used ACSA2-based cell sorting prior to single cell RNA sequencing (scRNAseq). We found a substantial population of ACSA2\n                      <jats:sup>+<\/jats:sup>\n                      cells exhibiting robust microglial gene expression signatures, suggesting contamination of purified astrocyte preparations.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>An intravenous antibody labeling strategy coupled with flow cytometry was used to determine whether contamination originated from circulating immune cells or microglia.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Contaminating cells were identified as CNS-resident microglia that express CD45, CD11b, and ACSA2.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>These findings caution against the usage of ACSA2 for astrocyte purification without exclusion markers to achieve high-purity astrocyte populations for downstream multi-omics analyses.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fncel.2026.1756677","type":"journal-article","created":{"date-parts":[[2026,2,18]],"date-time":"2026-02-18T05:33:37Z","timestamp":1771392817000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["ACSA2 is not astrocyte-specific: implications for cell sorting strategies in the rodent brain"],"prefix":"10.3389","volume":"20","author":[{"given":"Emerson","family":"Daniele","sequence":"first","affiliation":[{"name":"Institute of Medical Science, University of Toronto","place":["Toronto, ON, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Khelifi","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Medical Biochemistry and Pathology, Faculty of Medicine, Universit\u00e9 Laval, Qu\u00e9bec City","place":["QC, Canada"]},{"name":"Oncology Division, CHU de Qu\u00e9bec-Universit\u00e9 Laval Research Centre, Qu\u00e9bec City","place":["QC, Canada"]},{"name":"Universit\u00e9 Laval Cancer Research Centre, Qu\u00e9bec City","place":["QC, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Beretta","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, Hotchkiss Brain Institute, University of Calgary","place":["Calgary, AB, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boyan K.","family":"Tsankov","sequence":"additional","affiliation":[{"name":"Department of Immunology, University of Toronto","place":["Toronto, ON, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. 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Annie","family":"Bang","sequence":"additional","affiliation":[{"name":"Network Biology Collaborative Centre Flow Cytometry Facility, Sinai Health, Lunenfeld-Tanenbaum Research Institute","place":["Toronto, ON, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Beretta","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, Hotchkiss Brain Institute, University of Calgary","place":["Calgary, AB, Canada"]},{"name":"Alberta Children\u2019s Hospital Research Institute, University of Calgary","place":["Calgary, AB, Canada"]},{"name":"Hotchkiss Brain Institute, University of Calgary","place":["Calgary, AB, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dana J.","family":"Philpott","sequence":"additional","affiliation":[{"name":"Institute of Medical Science, University of Toronto","place":["Toronto, ON, Canada"]},{"name":"Department of Immunology, University of Toronto","place":["Toronto, ON, Canada"]},{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto","place":["Toronto, ON, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samer M.","family":"Hussein","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Medical Biochemistry and Pathology, Faculty of Medicine, Universit\u00e9 Laval, Qu\u00e9bec City","place":["QC, Canada"]},{"name":"Oncology Division, CHU de Qu\u00e9bec-Universit\u00e9 Laval Research Centre, Qu\u00e9bec City","place":["QC, Canada"]},{"name":"Universit\u00e9 Laval Cancer Research Centre, Qu\u00e9bec City","place":["QC, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Faiz","sequence":"additional","affiliation":[{"name":"Institute of Medical Science, University of Toronto","place":["Toronto, ON, Canada"]},{"name":"Department of Cell Biology and Anatomy, Hotchkiss Brain Institute, University of Calgary","place":["Calgary, AB, Canada"]},{"name":"Alberta Children\u2019s Hospital Research Institute, University of Calgary","place":["Calgary, AB, Canada"]},{"name":"Hotchkiss Brain Institute, University of Calgary","place":["Calgary, AB, Canada"]},{"name":"Department of Laboratory Medicine and Pathobiology, University of Toronto","place":["Toronto, ON, Canada"]},{"name":"Division of Anatomy, Department of Surgery, University of Toronto","place":["Toronto, ON, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,2,18]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"24385150","DOI":"10.1038\/nprot.2014.005","article-title":"Intravascular staining for discrimination of vascular and tissue leukocytes.","volume":"9","author":"Anderson","year":"2014","journal-title":"Nat. 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