Materials and Reagents

1. HEPES (Serva, catalog number: 25245 )
   
2. EDTA (Merck KGaA, catalog number: 108418 )
   
3. Dispase in HBSS (BD Bioscience, catalog number: 354235 )
   
4. Collagenase D (Roche Diagnostics, catalog number: 11088866001 )
   
5. DNAse I (Sigma-Aldrich, catalog number: DN25 )
   
6. 4% (v/v) FCS (PAN Biotech, catalog number: P30-3302 )
   
7. Ice-cold PBS (Gibco, catalog number: 14190-094 )
   
8. HBSS (Gibco, catalog number: 14175-053 )
   
9. Percoll (GE Healthcare, catalog number: 17-0891-01 )
   
10. 10x PBS (Ambion, catalog number: AM9624 )
    
11. RPMI (Gibco, catalog number: 21875-034)
    
12. Cell-dissociation buffer (CD buffer) (see Recipes)
    
13. Digestion solution (see Recipes)
    
14. 40% percoll (see Recipes)
    
15. 80% percoll (see Recipes)
    

Equipment

1. Incubator (New Brunswick Scientific, incubator shaker G25)
   
2. 15 ml tube (Greiner Bio-One, catalog number: 188161 )
   
3. Centrifuge (Thermo Scientific, model: Multifuge X1R )
   

Procedure