Media 105

Control cell lines were used as positive controls (MCF-7, SH-SY5Y) and to test differential response to treatment (SVOG-3e). MCF-7, an estrogen- and progesterone-receptor-positive human breast cancer cell line, was used as positive control for the anti-hormonal therapies [49 (link)]. Additionally, SH-SY5Y, a human neuroblastoma cell line, was used as positive control for chemotherapeutic agents [50 (link)]. KGN, a human AGCT cell line derived from a 63-year-old patient heterozygous for FOXL2 c.402C > G, was used as an additional AGCT cell line to evaluate potential AGCT response [51 (link)]. Finally, the immortalized human granulosa cell line SVOG-3e [52 (link)] was used to assess selective drug response in AGCT versus healthy granulosa cells. SH-SY5Y and MCF-7 were cultured in DMEM/F12 (Thermo Fischer Scientific) + 10% FBS and 1% Pen/Strep and RPMI 1640 (Thermo Fischer Scientific, Waltham, MA, USA) + 10% FBS and 1% Pen/Strep, respectively. KGN was cultured in conditions identical to AGCT-patient-derived cell lines. SVOG-3e was cultured in a 1:1 ratio of Media 199 and Media 105 (Sigma-Aldrich, St. Louis, MO, USA) with 5% FBS and 1% Pen/Strep.

The 184-hTERT cell lines were cultured at 37 °C, 5% CO₂, in serum-free mammary epithelial cell basal media (MEBM, Lonza), supplemented with mammary epithelial cell growth media single quots (Lonza), 5 μg/ml transferrin (Sigma), 1.25 M of isoproterenol (Sigma Aldrich). HCT116 cell lines were cultured at 37 °C, 5% CO₂, in McCoys 5A media (Sigma Aldrich) supplemented with 10% FBS (Sigma Aldrich). The TOV3133D and TOV3133G cell lines were cultured at 37 °C, 5% CO₂, in a 1:1 mix of media 199 (Sigma Aldrich) and media 105 (Sigma Aldrich) supplemented with 10% FBS.