Isolation of Primary Human Keratinocytes from Foreskin

Materials and Reagents

1. Foreskin
   
2. 0.05% Trypsin (Life Technologies, Invitrogen^TM, catalog number: 25300-054 )
   
3. 2x dispase (Life Technologies, Invitrogen^TM, catalog number: 17105-041 )
   
4. DMEM (Life Technologies, Invitrogen^TM, catalog number: 11965-092 )
   
5. Fetal bovine serum (FBS) (Life Technologies, Invitrogen^TM, catalog number: 16000-044 )
   
6. Defined keratinocyte serum-free medium (Life Technologies, Invitrogen^TM, catalog number: 10744-019 )
   
7. 154 medium (Life Technologies, Invitrogen^TM, catalog number: M-154-500 )
   
8. Penicillin and streptomycin (Life Technologies, Invitrogen^TM, catalog number: 10378-016 )
   
9. Phosphate buffered saline (PBS)
   
10. DMEM cultural medium (see Recipes)
    
11. 50: 50 medium (see Recipes)
    

Equipment

1. Centrifuges
   
2. Incubator (37 °C and 5% CO₂)
   
3. Sterile petridish
   
4. Cell culture hood
   
5. Cell culture dishes
   
6. Scissors
   
7. Forceps
   
8. 50 ml falcon tube
   
9. Disposable sterile cell culture strainer
   

Procedure