“…Brains were obtained from 13-day-old mouse embryos and placed in cold, sterile, phosphate-buffered saline (PBS). As previously described (Giorgi et al, 2015), the midbrain was dissected out under microscopic control and placed in a nutrient medium composed of F12 and Eagle's minimal essential medium (ratio1:1), supplemented with 2 mM glutamine, 10% of Nu-serum and 33.3 mM glucose. The tissue was mechanically dispersed, centrifuged at 800 g for 3 min, and suspended in the culture medium.…”