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In this application protocol, we describe a protocol to generate highly purified and viable oligodendrocytes from adult mouse or rat brain tissue. Brain tissue from mice or rats 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 are removed using the Debris Removal Solution and is followed by an subsequent removal of erythrocytes using the Red Blood Cell Removal Solution. The Anti-O4 MicroBeads are used to isolate oligodendrocytes 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|
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