dean g. tang laboratory
M.D. Anderson Cancer Center
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Other Protocols

 

 

Protocol for modified PrEBM (prostate primary epithelial cell culture)

To a 500 ml bottle of PrEBM (Clonetics CC-3165), add the following:

  • Insulin (Sigma I-6634) – 4 mg in 8 mls
    • 4 mg insulin/8 ml water. Add 40 ul 1N HCl. Filter sterilize.
  • Hydrocortisone (Sigma H-0396) – 250 ug
    • Make aliquots in water.
  • Epidermal Growth Factor (EGF) (Peprotech 100-15) – 5 ug
    • Make aliquots in water
  • Bovine pituitary extract (BPE) (Invitrogen 13028-014) - 25 ug
  • Penicillin/Streptomycin – 1/100
  • Fungizone (Gibco 15290-018) – 1/2000

Before plating cells, coat dishes with Vitrogen 100 (Cohesion FXP-019). Dilute 1/50 in water and coat either overnight at RT or 1 hour at 37° C. Wash 2X with water and let air-dry. Ready for use.