Cell Culture of Human Cancer Cells

Human OC cell lines, including OV90, OVCAR3, A2780, OVCAR8, SKOV3, and SW626, and human breast cancer cell line, HCC1937, were purchased from ATCC (Rockville, MD, USA) and cultured in MCDB 105 and Medium 199 (1:1), Dulbecco’s Modified Eagle Medium (DMEM), RPMI-1640, RPMI-1640, McCoy’5A, L-15, and RPMI-1640 media, respectively. MRC-5 human lung fibroblasts were obtained from the cell bank of the Chinese Academy of Sciences. Primary OC stromal fibroblasts were isolated and purified from fresh cancer tissues of OC patients as previously described^{50 (link)}, and baseline information is summarized in Supplementary Table 2, all participants provided written informed consent at recruitment, and the institutional ethics review committee of Tongji Hospital approved all study procedures for human subjects. MRC-5 cells were transformed into activated phenotype MRC5-CAFs by TGF-β1 (50 ng/ml). All fibroblasts were cultured in DMEM/F-12 (1:1) medium (Gibco). All the above growth media were supplemented with 1% penicillin/streptomycin (Thermo Scientific) and 10% fetal bovine serum (Gibco). All cells were cultured in a humidified atmosphere incubator with 5% CO₂ at 37 °C. Mycoplasma testing (Lonza) was performed regularly in our institution, and all cell lines were tested once after thawing or isolation and before other experiments.