Culturing Human Prostate Epithelial Cell Lines

Immortalized 957E/hTERT and 957E/hTERT-AR human prostate epithelial cells were generously provided by John T. Isaacs lab (Johns Hopkins, Baltimore, MD, USA) [9 (link),10 (link)]. The prostate epithelial cell line RWPE-1 was purchased from the American Type Culture Collection (ATCC, Cat. No. CRL-11609). The cells, 957E/hTERT, 957E/hTERT-AR and RWPE-1, were grown in Keratinocyte-SFM (serum-free medium) supplemented with bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF) supplied as a kit (Cat. No. 17005-042, Thermo Fisher Scientific, Waltham, MA, USA), and 1% penicillin-streptomycin. Primary prostate epithelial cells (PrEC) were purchased from ATCC, Manassas, VA, USA (Cat. No. PCS-440-010) and were grown in prostate epithelial cell basal medium (ATCC, PCS-440-030), supplemented with prostate epithelial cell growth kit contents (ATCC, PCS-440-040). The prostate epithelial cell line EP156T was grown in MCDB153 medium (Biological Ind. Ltd., Beit Haemek, Israel), supplemented with 1% fetal calf serum (FCS), growth factors and antibiotics as described [3 (link),5 (link)]. Complete growth media were changed every 2–3 days.