Splitting of Rat Astrocytes
16.2.1. Astrocytes are split into 6 x uncoated T75 flasks.
16.2.2 Adherent astrocytes are washed with PBS once.
16.2.3. Add 10mL of Accutase per T75 flask and incubate the dish at 37°C for approximately 10-20 mins.
16.2.4. Check regularly for cell lifting.
16.2.5. Carefully transfer all of the dish content into a pre-warmed falcon.
16.2.6. Centrifuge at 350g for 5 mins.
16.2.7. While spinning, add fresh AMM to the new T75 flasks.
16.2.8. Aspirate media from cell pellet in spinning falcon and replace with AMM, slowly and gently resuspending the pellet.
16.2.9. Transfer an equal amount of cell suspension into previously prepared T75 flasks and swirl plate gently in a figure 8 motion.
16.2.10. Expand again for another 2 to 3 weeks till confluence.