Isolating Murine Primary Oligodendrocytes Using Immunomagnetic Beads
Transcription
Take a neural cell suspension, including immature oligodendrocytes and primary neuronal cells, from a mouse pup.
Resuspend the cells in a magnetic cell sorting buffer to avoid cell clumping.
Add engineered magnetic microbeads carrying anti-O4 antibodies on their surface. Agitate the solution to ensure uniform mixing.
Incubate to enable microbeads to bind with sulfated galactolipid, an O4 antigen on oligodendrocytes, while other cells remain unbound or loosely bound.
Wash the mix with additional cell sorting buffer.
Centrifuge and discard the supernatant, then resuspend the cells.
Next, pass the bead-bound cells into a magnetic separation column equipped with a filter that removes the larger cell clumps.
During the run, under a magnetic field, the bead-bound complexes move toward the magnet, attaching to the sides.
Wash to remove unbound and loosely bound cells.
Retrieve the column from the separator and wash it with a proliferation medium to collect the primary oligodendrocytes for future applications.
