Materials and Reagents

1. Poly-D-lysine (Sigma-Aldrich, catalog number: P7280 )
   
2. MEM (Life Technologies, Gibco^®, catalog number: 11090-08 )
   
3. D-Glucose
   
4. Sterile water
   
5. Sterile PBS
   
6. Trypan blue dye
   
7. Heat-inactivated fetal bovine serum (FBS) (Life Technologies, Gibco^®, catalog number: 16000-044 )
   
8. Heat-inactivated horse serum (HS) (Life Technologies, Gibco^®, catalog number: 26050-088 )
   
9. None essential nonessential amino acids (Life Technologies, Gibco^®, catalog number: 11140-050 ) (100 ml)
   
10. Sodium pyruvate (Sigma-Aldrich, catalog number: S8636 ) (100 ml)
    
11. 200 mM L-glutamine (Life Technologies, Gibco^®, catalog number: 25030-081 ) (100 ml)
    
12. Penicillin/streptomycin (Sigma-Aldrich, catalog number: P0781 ) (100 ml)
    
13. Poly-D-lysine stock solution (see Recipes)
    
14. Maintenance culture medium (see Recipes)
    
15. Treatment medium (see Recipes)
    

Equipment

1. Cell culture incubator
   
2. Standard benchtop centrifuges
   
3. Hemocytometer
   
4. Dissection microscope
   
5. Scissors and forceps
   
6. Sterile filter (0.2 µm)
   
7. Foil
   
8. 24-well plates
   
9. Laminar hood
   
10. 50-ml tube
    
11. 10-ml pipet
    

Procedure