Androgen Production in H295R Cells

Human adrenocortical-carcinoma cells (H295R cells) were used as an in vitro cell culture model for androgen production. These cells are commonly used in studies of steroidogenesis and androgen biosynthesis pathways [21 (link)–23 (link)]. H295R cells were purchased from ATCC (Manassas, VA, USA, cat. no. ATCC® CRL-2128™) and cultured per the recommended guidelines. Briefly, H295R cells were cultured in flasks pre-coated with extracellular matrix (Gibco, USA, cat. no. S-006-100) with DMEM/F12 (Gibco, cat. no. 21041025) and 2.5% Nu-Serum (Corning, USA). The cells were subcultured at a ratio of 1:3 to 1:4 and culture media were changed twice a week.