To begin delineating the cellular and molecular events that are important in ovarian carcinogenesis, we have developed a simple and rapid method for the establishment of primary cultures derived from benign tumors, malignant tumors, and ascites of the ovary that are representative of the original clinical material from which they are derived. From 23 ovarian epithelial ascites collected, 13 were successfully established in culture and cells survived an average of 7 to 8 passages. From 65 solid epithelial ovarian tumors (benign and malignant) 36 were cultured for an average of 6 passages for cultures derived from benign tumors and 11 or 12 passages in the case of malignant tumors. Cells were scored as epithelial in nature by morphology and histochemical analysis using anti-cytokeratin antibodies. Cultures, especially those derived from solid tumors, sometimes displayed fibroblastic-like contamination which was quickly resolved. We include limited molecular analyses both to characterize the origin of the populations we have established as well as to demonstrate the usefulness of these cultures in molecular studies.