Isolating Immune Cells from Mouse Choroid Plexuses
Transcription
Position a mouse's brain dorsal side up.
Under a microscope, separate the cerebral hemispheres. Pull away the cortex and then the hippocampus to expose the lateral ventricles and the third ventricle.
Collect two lateral choroid plexuses. These are immune-cell-rich brain structures that line the lateral ventricles like a long veil.
Collect the third ventricle choroid plexus, which is a short structure with a granular surface.
Detach the cerebellum to expose the fourth ventricle.
Collect the fourth ventricle choroid plexus, which is a long globular structure with a granular surface.
Place all the choroid plexuses in a tube with buffered saline to maintain tissue integrity.
Add a digestive enzyme solution and incubate with agitation.
The enzyme breaks down the extracellular matrix and loosens the cells.
Add a suitable buffer to inactivate the enzyme.
Centrifuge to settle the cells. Discard the supernatant and resuspend the cells in a fresh buffer for further analysis.
