adult brain dissociation kit  (Miltenyi Biotec)

Journal: Cells

Article Title: Astro-Versus Microglia-Enriched Transcriptomes from Aged Atxn2 -CAG100-Knockin Mice Suggest Underlying Pathology of RNA Processing at Ribosomes, and Possibly at U-Bodies

doi: 10.3390/cells15080699

Figure Lengend Snippet: Schematic workflow for adult brain dissociation and sequential isolation of cell types (microglia, astrocytes, and neurons). Tissue is harvested rapidly, placed in ice-cold DPBS with Ca 2+ and Mg 2+ , and cut into small pieces, then transferred to C-tubes and dissociated by a combination of enzymatic buffer solution containing papain with gentle mechanical dissociation, using a gentleMACS Octo Dissociator with heaters. Gradient centrifugation forms a compact ring containing debris (dead cells and myelin). After debris removal, red blood cells are lysed, and the dissociated cells are magnetically labeled with anti-CD11b MicroBeads against microglia. The labeled cells are passed through LS columns twice and placed on a magnetic stand. CD11b-positively selected cells remain in the column and are eluted with a plunger into a fresh tube. The negative flow-through is processed to isolate astrocytes labeled with anti-ACSA2 MicroBeads, and the process is repeated. For neuron isolation, the negative flow-through from astrocyte extraction is labeled with a non-neuronal biotin cocktail and anti-biotin MicroBeads, and the subsequent negative flow-through contains the neuron population. The arrows show the workflow handling of the cell suspensions. Image created in BioRender, Reddy, A. (2026) https://BioRender.com/mj9rqid , last accessed on 2 March 2026 and adapted from .

Article Snippet: The subsequent protocol is adapted from Miltenyi adult brain dissociation (Miltenyi, #130-107-677, Miltenyi Biotech, Bergisch Gladbach, Germany): adult neuron isolation, mouse (Miltenyi, #130-126-603); Anti-ACSA2 MicroBead Kit, mouse (Miltenyi, #130-097-679); and CD11b (Microglia) MicroBeads, human and mouse (Miltenyi, #130-093-636)—see [ , ].

Techniques: Isolation, Gentle, Gradient Centrifugation, Labeling, Extraction