2.3. Neurosphere culture myelin depletion

Percoll Plus solution (23%, Millipore‐Sigma, GE17‐5445‐01) dissolved in 1X sterile phosphate‐buffered saline (PBS) was added to primary enzymatically digested tissue (brain and spinal cord) and triturated until tissue was homogenized in the solution. The solution was then centrifuged for 15 minutes at 1800 RPM. Myelin supernatant was removed and myelin depleted tissue was washed with serum‐free media (37°C). The resuspended pellet was triturated until homogenized and centrifuged at 1500 RPM for 3 minutes. Supernatant was removed and cells were counted and plated at clonal density in the neurosphere assay as described. 12