Establishment of Ovarian Cancer Cell Lines

In total, six cell lines were derived from samples from six patients, 866, 2978, 3041, 3291, 4453 and 4485. All cell lines were maintained in a low oxygen condition of 7% O₂, and 5% CO₂ and grown in complete OSE medium, which includes OSE medium (Wisent, St-Bruno, QC), 10% FBS, 0.5 μg/mL amphotericin B (Wisent) and 50 μg/mL gentamicin (Gibco®, Life Technologies Inc., Burlington, ON). The solid ovarian tumor (TOV)-derived cell lines (TOV2978G, TOV3041G, TOV3291G) were established using the scrape method as previously described [11 (link), 76 (link)]. Briefly, tumor tissue was scraped into a 100 mm plate with complete OSE medium and maintained for 40 days with the medium replaced weekly. Cells were passaged at near confluence, and were considered immortal when passaged over 50 times. The OV cell lines (OV866(2), OV4453, OV4485) were established from the cellular fraction of ascites collected by centrifugation [11 (link), 76 (link)]. The cell lines derived from ascites cells were maintained as above for the TOV derived cell lines. Although to date each cell line has reached at least 90-100 passages, most assays were conducted on cell lines between passage 60 and 80.