To begin, add 5 milliliters of 10 micrograms per milliliter Poly-L-ornithine hydrobromide to each T75 flask. Incubate at 37 degrees Celsius in a 5%carbon dioxide incubator for at least one hour. After decapitating the mouse, gently hold the head and create a midline incision in the cranium, starting from the neck to the nose.
Peel the skull back with tissue forceps to reveal the brain. To scoop the brain out, insert the tissue forceps under the brain, and lift the brain out from the head. After placing the brain into a Petri dish containing 20 milliliters of Leibovitz's L-15 media under a dissection microscope, using a pair of micro-forceps, carefully remove the brainstem and the meninges.
Collect the brains in a 50-milliliter conical tube containing 5 milliliters of Leibovitz's L15 media.
