title: Establishment of primary cultures from ovarian tumor tissue and ascites fluid.
abstract: We have refined the technique for isolating and propagating cultures of primary epithelial ovarian cancer (EOC) cells derived from solid tumors and ascites. Both protocols involve a simple yet rapid method for the growth and propagation of EOC tumor and ascites cells in a basal culture medium without the addition of growth factors. Isolation of tumor EOC cells involves the mechanical disruption of the tumor tissue with the help of a cell scraper, while ascites-derived EOC cells are mixed with growth medium and placed directly into culture with very little manipulation. We further describe a partial trypsinization method to eliminate fibroblast contamination from primary EOC cells derived from solid tumors. These methods allow for the direct application of many molecular, cellular, and functional analyses within a few weeks of initial isolation, with the added potential of retrospective analyses of archived cells and tissues. Thus, we have included steps for long-term cryopreservation of early-passage EOC cells. Initial isolation of EOC cells can be completed within 1 h, and primary cells are further expanded in culture for several weeks.
authors.affiliation: Campbell Family Cancer Research Institute, Ontario Cancer Institute, University Health Network, Toronto, ON, Canada.
authors.forename: Brigitte L
authors.initials: BL
authors.lastname: Thériault
authors.forename: Lise
authors.initials: L
authors.lastname: Portelance
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Mes-Masson
authors.forename: Mark W
authors.initials: MW
authors.lastname: Nachtigal
mesh_terms: D001202:Ascitic Fluid / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology*; D061251:Primary Cell Culture / Q000379:methods*