ARPE-19 Cell Culture Protocol

Human ARPE-19 cells (ATCC, Manassas, VA, United States, Cat. # CRL-2302) were maintained in Dulbecco’s modified eagle medium/Nutrient Mixture F-12 (DMEM/F-12) (Gibco; Grand Island, NY, Cat. #11330-032) supplemented in 10% fetal bovine serum (FBS) (Gibco; Grand Island, NY Cat#10082-147) and 1% penicillin/streptomycin (Gibco; Grand Island, NY Cat.# 15070-063) at 37°C with 5% CO as previously described (Bullock et al., 2021 (link)). For assays described below, a total of 0.2 × 10⁶ cells in 0.5 ml were plated per well of a 24-well plate and incubated for 72 hours in DMEM/F12 and 1% penicillin-streptomycin and no FBS before cell treatment. ARPE-19 cells were authenticated by Bio-Synthesis, Inc. (Lewisville, TX) at passage 27. ARPE-19 cells in passage numbers 27–32 were used for all experiments.