This application protocol describes a protocol to generate highly purified and viable astrocytes from adult mouse brain tissue. Brain tissue from mice older than P7 is dissociated into single-cell suspensions using the Adult Brain Dissociation Kit. The extracellular matrix is enzymatically digested using the kit components, while the gentleMACS™ Dissociator with Heaters is used for the mechanical dissociation steps during the on-instrument enzyme incubation. After the dissociation, the myelin and cell debris is removed using the Debris Removal Solution and is followed by a subsequent removal of erythrocytes using the Red Blood Cell Removal Solution (10×). The Anti-ACSA-2 MicroBead Kit is used to isolate astrocytes from the single-cell suspension.
|Enzyme mix 1||Enzyme mix 2|
|Debris Removal Solution||D-PBS||Overlay (D-PBS)|
|900 µL||3100 µL||4 mL|
|1800 µL||6200 µL||4 mL|
|Column||Max. number of labeled cells||Max. number of total cells||Selector|
VarioMACS, SuperMACS II
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