Isolation of Cells from Sensory Ganglia
Transcription
Take prewashed sensory ganglia in a tube.
Add a dissociation solution containing a mixture of digestive enzymes and incubate.
These enzymes break down the extracellular matrix, releasing individual cells.
After incubation, remove the dissociation solution.
Wash the ganglia with a phosphate buffer, and add a fresh dissociation solution.
Pipette the ganglia up and down to dissociate the cells.
Pass the suspension through a cell strainer to remove any clumps.
Collect any remaining cells from the bottom of the strainer and put them into the tube.
Layer the cell suspension over a density gradient tube and centrifuge.
Based on the differences in shape and mass, cells migrate to the lower layer, while cellular debris remains in the upper layer.
Discard the upper layer.
Add fresh medium to the remaining cell suspension and centrifuge.
Discard the supernatant and store the sensory ganglia cells for further use.
