Culturing Docetaxel-Resistant Prostate Cancer PC3 Cells

Prostate cancer cell line PC3 was acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA, Cat# ATCC-CRL-1435) and cultured in RPMI-1640 medium (Genesee Scientific, San Diego, CA, USA, Cat# 25-506), supplemented with 10% (v/v) fetal bovine serum (Genesee Scientific, San Diego, CA, USA, Cat# 25-514), penicillin/streptomycin (Fisher Scientific, Pittsburgh, PA, USA, Cat# 30-002-CI), and normocin 1G (Fisher Scientific, Cat# NC9390718). Cells were grown under 5% CO₂ at 37 °C. A docetaxel-resistant PC3 cell line (PC3-DR) was developed as indicated previously [50 (link)] and maintained in the presence of 10 nM docetaxel (LC Laboratories, Woburn, MA, USA, Cat# D-1000). This PC3-DR cell line was recently shown by our group to upregulate DFS70/LEDGFp75 and members of its protein interactome and is an excellent model for evaluating the immunoreactivity of antibodies to these proteins [50 (link)]. Short tandem repeat (STR) service provided by ATCC (Cat# ATCC-135-XV) was used to authenticate the PC3 and PC3-DR cell lines. Mycoplasma testing was conducted at least twice a year using the MycoAlert^TM Mycoplasma Detection Kit (Lonza, Walkersville, MD, USA, Cat# LT07-218). Cells that were found contaminated were discarded. Cell lines were grown until approximately 20 passages.