Human prostate tumor cell line CW22Rv1 (22Rv1) was purchased from ATCC and grown in RPMI-1640 supplemented with either 10% fetal bovine serum (FBS) or 3% charcoal-stripped FBS (for palmitic acid and cholesterol treatment where indicated), and 1% penicillin/streptomycin (all components from Thermo Fisher). Human androgen-refractory prostate cancer cells of the ARCaPM cell line were gifted to us from Leland Chung (Cedars-Sinai Medical Center), and grown in DMEM, supplemented with 5% FBS and 1% penicillin/streptomycin [24 (link)].
Human primary fibroblasts were grown from prostatectomy specimens at Cedars-Sinai Medical Center or the Greater Los Angeles Veterans Affairs under their respective institutional review boards [42 (link)]. The tumor-inductive status of fibroblasts was determined by tissue recombination with BPH1 (a non-tumorigenic human prostate epithelial cell line, as previously described [32 (link)]). CAF was cultured in DMEM/F12 supplemented with 5% FBS, 5% Nu-Serum, 1% penicillin/streptomycin, 10−9 M testosterone (Sigma-Aldrich), and 4 μg/mL insulin (12585014, Fisher Scientific). All cells were grown in a humidified incubator at 37 °C with 5% CO2. All cells were tested for mycoplasma (LT07118, Lonza, Rockland, ME, USA) every 1 month and were negative.
Human primary fibroblasts were grown from prostatectomy specimens at Cedars-Sinai Medical Center or the Greater Los Angeles Veterans Affairs under their respective institutional review boards [42 (link)]. The tumor-inductive status of fibroblasts was determined by tissue recombination with BPH1 (a non-tumorigenic human prostate epithelial cell line, as previously described [32 (link)]). CAF was cultured in DMEM/F12 supplemented with 5% FBS, 5% Nu-Serum, 1% penicillin/streptomycin, 10−9 M testosterone (Sigma-Aldrich), and 4 μg/mL insulin (12585014, Fisher Scientific). All cells were grown in a humidified incubator at 37 °C with 5% CO2. All cells were tested for mycoplasma (LT07118, Lonza, Rockland, ME, USA) every 1 month and were negative.