Culturing Mouse and Human Prostate Cancer Cells

Mouse MyC-CaP (53 (link)) and luciferase-expressing MyC-CaP (MyC-CaP/GFP-Luc) (54 (link)) PCa cells were grown in Dulbecco’s minimal essential medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin (100 U/ml) and streptomycin (100 μg/ml; 1X Pen/Strep). Both cell lines were confirmed to be negative for mycoplasma, lymphocytic choriomeningitis virus, lactate dehydrogenase virus, mouse hepatitis virus, and mouse parvovirus by IDEXX Bioanalytics. Human C4-2B PCa cells were purchased from American Type Culture Collection and were grown on 0.01% poly-lysine–coated tissue culture plates in RPMI 1640 supplemented with 10% FBS and 1X Pen/Strep. All cells were maintained in a sterile incubator at 37°C and 5% CO₂. For serum starvation, cells were cultured in growth media with 0.1% FBS. For pervanadate stimulation, cells were treated with 200 μM pervanadate for 15 min at 37°C before lysis.