Isolating Vestibular Ganglion Neurons From a Mouse Inner Ear

Take one-half of a mouse pup's head and remove the brain.

Cut the top of the skull and excise the surrounding tissue to visualize the otic capsule, a bony outer layer protecting the inner ear structures.

Transfer the tissue to a culture plate containing a buffered medium.

Separate the superior and inferior vestibular ganglia, which contain the vestibular nerve cell bodies.

Shave off the bony ridge and harvest the underlying superior ganglion.

Transfer the superior ganglion to a fresh plate containing the buffered medium.

Remove adherent tissues from the surface to clean the isolated ganglion.

Place the ganglion in a solution containing proteolytic enzymes.

The enzymes degrade the tissue's extracellular matrix, loosening the cells.

Transfer the ganglion to a polymer-coated glass-bottom dish containing culture medium and mechanically dissociate the tissue to release the cells.

Incubate, allowing the cells to adhere and establish a vestibular ganglion neuron culture.

Scoop out the brain of the euthanized mouse using closed surgical scissors. Sever and remove the cranial nerve to completely detach the brain from the skull. Starting at the back of the head, pull the clear membranous mat