Culturing Murine Spiral Ganglion Neuron Explants on Multielectrode Arrays
Transkript
Separate the organ of Corti from the spiral ganglion in modiolus by holding the basal portion of the spiral ganglion and the organ of Corti with forceps, and slowly unwinding the organ of Corti from base to apex. Then, cut the lateral explants from the spiral ganglion using forceps or fine microdissection scissors or knives.
Now, transfer the spiral ganglion explants and the organ of Corti onto the MEA. Then, place SG explants over the electrode area and the organ of Corti, approximately 5 millimeters away from the electrode area. Place the explants and the organ of Corti onto the MEAs while avoiding damage to the tissue. Afterward, place the MEAs carefully in the incubator and culture at 37 degrees Celsius and 5% CO2.
The next day, visually inspect the explants for their attachment to the MEAs. Then, add 100 microliters of culture medium containing 10% FBS and BDNF daily for five consecutive days. On day 6, add 2 milliliters of culture medium, containing 10% FBS and 5 nanograms per milliliter BDNF, and culture the tissue for an additional 13 days.
