Lncap and pc 3 human prostate cancer cell lines

LNCaP and PC‐3 human prostate cancer cell lines were purchased from the American Type Culture Collection (Manassas, Virginia, USA). Human SaOS‐2 osteosarcoma cells were purchased from the European Collection of Cell Cultures (Salisbury, UK). The LNCaP‐SF androgen‐independent LNCaP cell line was established in our laboratory by long‐term subculture of the parental LNCaP cells in RPMI (Thermo Fisher Scientific, Waltham, Massachusetts, USA) with 5% charcoal‐stripped FBS (CCS; Hyclone, Logan, Utah, USA) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, California, USA).17 Bone‐derived stromal cells (BDSC) and bone metastasis stromal cells (BmetSC) were obtained from normal eleventh ribs and left thigh bones with prostate cancer metastases as previously described.7 The bone stromal cells were cultured in RPMI supplemented with 1% penicillin/streptomycin and 10% FBS (Sigma‐Aldrich, St. Louis, Missouri, USA). LNCaP and PC‐3 cells were cultured in RPMI supplemented with 5% FBS and 1% penicillin/streptomycin. SaOS‐2 cells were cultured in RPMI supplemented with 1% penicillin/streptomycin and 5% FBS. All cells were kept at 37°C in a humidified atmosphere with 5% CO₂.