Establishment and Characterization of Ovarian Cancer Cell Lines

The OCI cell lines and OCMI medium are available from the Live Tumor Culture Core (LTCC) at Sylvester Comprehensive Cancer Center (SCCC), Miller School of Medicine, University of Miami http://sylvester.org/shared-resources/Live-Tumor-Culture-Core In order to establish the cell lines, fresh tumor tissue fragments are minced and plated on Primaria (BD Biosciences) plates before and after digestion with 1 mg/ml collagenase (Roche). The tumor cells are cultured in nutrient medium described previously ^{16 (link)}, supplemented with insulin, hydrocortisone, EGF, cholera toxin, and serum as described in Supplementary Methods. We refer to this version of the medium as Ovarian Carcinoma Modified Ince medium (OCMI). This formulation is supplemented with 17β-estradiol for endometrioid and mucinous tumors (OCMIe). The papillary serous, clear cell, dysgerminoma, and carcinosarcoma tumors are initially cultured in 5% CO₂ and regular O₂ at 37 C as monolayers attached to Primaria culture plates. The endometrioid and mucinous tumors are initially cultured in 5% CO₂ and low O₂ (5%) at 37 C as monolayers attached to Primaria culture plates. Adjustments in O₂ and plates are made for individual cell lines as necessary (Supplementary Table 3). The tumor cells are passaged at a ~ 1:3 ratio once a week and plated into a new flask at approximately 1×10⁴ cells/cm². During the initial weeks of culture, (~1–5) the plates are treated with diluted trypsin first, in order to deplete stromal cells. The remaining cells that are still attached to the culture plate are treated with 0.25% trypsin for sub-culturing. In general, tumor cultures are free of stromal and normal cell types within 4–6 passages. The SOC cell lines were STR validated and cultured as per the instructions of the vendor. OCI lines will be available from the Ince laboratory upon publication. All study procedures were approved by the Institutional Review Boards of the University of Miami, Brigham & Women’s and Massachusetts General Hospitals to collect discarded tissues with written consent from all patients. See Supplementary Note 1 for further details of culture methods.