Fetuin

PC346C (p25) was generated in our lab as described previously (25 (link), 26 ) and maintained in prostate growth medium (PGM) consisting of DMEM-F12, supplemented with 2% FCS (both from Lonza, Breda, Netherlands), 1% insulin-transferrin-selenium (Gibco), 100 ng/ml fibronectin (Harbor Bio-Products, Tebu-bio, Netherlands), 20 μg/ml fetuin (ICN Biomedicals, Zoetermeer, Netherlands), 0.01% BSA, 10 ng/ml epidermal growth factor, 50 ng/ml cholera toxin, 0.1 mM phosphoethanolamine, 0.6 ng/ml triiodothyronine, 500 ng/ml dexamethasone (all from Sigma-Aldrich), 0.1 nM R1881 (Sigma), and penicillin/streptomycin antibiotics (100 U/ml penicillin, 100 μg/ml streptomycin; Cambrex BioWhittaker). VCaP (p30) (20 ) (kindly provided by Dr. K.J. Pienta, Baltimore, MD, USA) and DuCaP (p52) (21 ) (kindly provided by Dr. J.A. Schalken, Nijmegen, Netherlands) were maintained in RPMI1640 (Lonza), supplemented with 10% FCS and penicillin/streptomycin antibiotics. LAPC4 (p49) (22 (link)) was kindly provided by Organon (Oss, Netherlands) and maintained in Iscove's Modified Dulbecco's Medium (IMDM) (Lonza), supplemented with 7.5% FCS and 10 nM R1881. Cell line authentication was performed by short tandem repeat analysis by the Promega PowerPlex 16 kit.

Human PCa cells PC3, LNCaP and 22RV1 were maintained in RPMI-1640 medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 100 ug/ml penicillin/streptomycin. VCAP and LAPC4 Cell lines were maintained in Dulbecco's Modified Eagle Medium (DMEM, Invitrogen) supplemented with 10% FBS and 1% penicillin/streptomycin (Invitrogen). PC346C cells were cultured in Dulbecco's modified Eagle's medium-Ham's F-12 medium ( Invitrogen) supplemented with 0.1 nm R1881, 2% fetal calf serum (PAN Biotech), 1% insulin-transferrin-selenium (Invitrogen), 0.01% BSA (Roche), 10 ng/ml epidermal growth factor (Sigma-Aldrich), 1% penicillin/streptomycin, 100 ng/ml fibronectin (Harbor Bio-Products), 20 μg/ml fetuin (ICN Biomedicals), 50 ng/ml cholera toxin (Sigma-Aldrich), 0.1 mm phosphoethanolamine (Sigma-Aldrich), and 0.6 ng/ml triiodothyronine (Sigma-Aldrich. PC346C cells were obtained from Martin Tenniswood, University of Albany. All other cell lines were obtained from the American Type Culture Collection. Cell were obtained between 2001 and 2012, expanded, frozen and stored as stocks in liquid nitrogen. Cells were discarded after 10 passages after revival. Mycoplasma testing was carried out monthly. All cell lines are authenticated by STR analysis at MD Anderson Cancer Center Characterized Cell Line Core Facility.

Human PCa cells PC3, LNCaP, DU145 and 22RV1 were maintained in RPMI-1640 medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 100 ug/ml penicillin/streptomycin. VCAP and LAPC4 Cell lines were maintained in Dulbecco's Modified Eagle Medium (DMEM, Invitrogen) supplemented with 10% FBS and 1% penicillin/streptomycin (Invitrogen). PC346C cells were cultured in Dulbecco's modified Eagle's medium-Ham's F-12 medium ( Invitrogen) supplemented with 0.1 nm R1881, 2% fetal calf serum (PAN Biotech), 1% insulin-transferrin-selenium (Invitrogen), 0.01% BSA (Roche), 10 ng/ml epidermal growth factor (Sigma-Aldrich), 1% penicillin/streptomycin, 100 ng/ml fibronectin (Harbor Bio-Products), 20 μg/ml fetuin (ICN Biomedicals), 50 ng/ml cholera toxin (Sigma-Aldrich), 0.1 mm phosphoethanolamine (Sigma-Aldrich), and 0.6 ng/ml triiodothyronine (Sigma-Aldrich). All cell lines are authenticated by STR analysis at MD Anderson Cancer Center Characterized Cell Line Core Facility.