Cultivation of Human Ovarian Cell Lines

We maintained human serous ovarian cancer cell lines including OVCA420, OVCA433, OVCA429, TYK-nu, SKOV8, CAOV3, DOV13, HEYA8, A2780 and SKOV3 [8 (link)], in RPMI 1640 (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS) and 100 IU/ml penicillin and 100 μg/ml streptomycin (Nacali Tesque). The immortalized human ovarian surface epithelial cell line, HOSE-E7 [20 (link)], kindly provided by Dr. Katabuchi at Kumamoto University, was cultured in DMEM/F12 (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% FBS and 100 IU/ml penicillin and 100 μg/ml streptomycin (Nacalai Tesque). 293FT cells were purchased from Thermo Fisher Scientific, and cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% FBS and 100 IU/ml penicillin and 100 μg/ml streptomycin. All cells were seeded into Cellstars® tissue culture plates (Greiner, Frickenhausen, Germany) in a humidified incubator containing 5% CO₂ at 37°C. All cell lines were regularly tested for mycoplasma.
As an inhibitor of the p38 MAPK pathway, we used SB203580 (Sigma-Aldrich, St.Louis, USA). To inhibit the ERK1/2 pathway, we used GSK1120212 (Sigma-Aldrich).