Enhanced Diagnostic Tools
Cell culture and treatment Benign human prostate epithelial cells (PrEC) were purchased from Lonza Walkersville Inc. (Walkersville, MA). LNCaP (ATCC CRL-1740) and PC3 (ATCC CRL-1435) cell lines were obtained from the American Type Culture Collection (Manassas, VA). Recent cultures of PC3 and LNCaP cells were analyzed using single-locus short tandem repeats (STR) DNA typing by Biosynthesis Inc. (Lewisville, TX) to authenticate cell identity. No contamination or misidentification was detected (data not shown). C4-2B cells were purchased from ViroMed Laboratories (Minnetonka, MN). PrEC cells were grown in serum-free prostate epithelial growth media (PrEGM) (Lonza/Cambrex). LNCaP, C4-2B and PC3 cells were maintained in RPMI 1640 supplemented with 5% fetal bovine serum and 1% antibiotic-antimycotic (Life Technologies, Inc., Rockville, MD) at 37°C in a humidified atmosphere of 95% air and 5% CO2, and cells grown less than 30 passages were used. In experiments designed to assess the effects of androgen treatment, cells were seeded in androgen-depleted media containing 5% Charcoal/Dextran treated FBS and 1% antibiotic-antimycotic for 24 h before R1881 treatment. R1881 concentration used in this study was 1 nM, an optimal dose stimulating growth of LNCaP cells. Vehicle was 0.001% ethanol.
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