Prostate Cancer Cell Culture Protocol

LNCaP cells were purchased from ATCC and cultured in RPMI1640 supplemented with 10% FBS (Hyclone, Logan, UT or Invitrogen, Carlsbad, CA, USA). LNCaP-TGF-β1(a) cells overexpressing an HA-tagged constitutively activated TGF-β1 ligand, HA-TGF-β1(a), and control LNCaP (LNCaP-Ctrl) cells were generated as previously described [37 (link)]. Human prostate stromal cell lines HPS19I, HTS33B, and HPS33Q were prepared following a previously described protocol and cultured in Bfs medium: DMEM supplemented with 5% FBS (Hyclone or Invitrogen), 5% Nu serum (Collaborative Research, Bedford, MA), 0.5 μg/mL testosterone, 5 μg/mL insulin, 100 units/mL penicillin, and 100 μg/mL streptomycin (Sigma, St. Louis, MO, USA) [5 (link), 38 (link)].