Ovarian Cancer Cell Line Establishment

The human ovarian epithelial cancer cells, A2780CP20, HEYA8, and HEYA8.MDR, were provided by Dr. Anil K. Sood (MD Anderson Cancer Center, Houston, TX, USA) and have been described elsewhere [78 (link)]. A2780 was purchased from the European Collection of Cell Cultures (ECACC, Porton Down, Salisbury, UK), and OV90 and OVCAR3 from the American Type Culture Collection (ATCC, Manassas, VA, USA)—both of which provide authenticated cell lines. OV90CIS and OVCAR3CIS cells were generated by exposing OV90 and OVCAR3 to increasing concentrations of cisplatin (CIS; Sigma-Aldrich, St. Louis, MO, USA). Cells were maintained in RPMI1640 (A2780, A2780CP20, HEYA8, and HEYA8.MDR; HyClone, GE Healthcare Life Sciences, Logan, UT, USA), RPMI1640 + insulin (0.01 mg/mL, OVCAR3 and OVCAR3CIS; Sigma) or M199/MCDB-105 (OV90 and OV90CIS; Gibco, Thermo Fisher Scientific, Grand Island, NY/Sigma) medium supplemented with 10% fetal bovine serum (FBS; HyClone) and 0.1% antibiotic/antimycotic solution (HyClone) at 37 °C in 5% CO₂ and 95% air. All cell lines were screened for mycoplasma using the LookOut^® Mycoplasma PCR detection kit (Sigma), and authenticated by Promega (Madison, WI, USA) and ATCC using Short Tandem Repeat (STR) analysis. In vitro assays were performed at 70%–85% cell density.