Generation of Microglia Cultures and Mixed Glial Culture

Materials and Reagents

1. Poly-D-lysine (Sigma-Aldrich, catalog number: P7280 )
   
2. DMEM/F12 (Life Technologies, Invitrogen^TM, catalog number: 11330-032 )
   
3. D-Glucose
   
4. Sterile water
   
5. Sterile PBS
   
6. Heat-inactivated fetal bovine serum (FBS) (Life Technologies, Gibco^®, catalog number: 16000-044 )
   
7. Non-essential amino acids (100 ml) (Life Technologies, Gibco^®, catalog number: 11140-050 )
   
8. Sodium pyruvate (100 ml) (Sigma-Aldrich, catalog number: S8636 )
   
9. 200 mM L-glutamine (100 ml) (Life Technologies, Gibco^®, catalog number: 25030-081 )
   
10. Penicillin/streptomycin (100 ml) (Sigma-Aldrich, catalog number: P0781 )
    
11. DMEM/F12-based culture medium (see Recipes)
    
12. Poly-D-lysine stock solution (see Recipes)
    
13. Treatment medium (see Recipes)
    

Equipment

1. Cell culture incubator
   
2. Centrifuges
   
3. Dissection microscope
   
4. Scissors and forceps
   
5. T175 flasks
   
6. 70 µm nylon filter (sterile)
   
7. 50 ml tube
   
8. Sterile filter (0.2 µm)
   
9. Foil
   
10. Laminar hood
    

Procedure