Search

Objective: Ovarian cancer is a poor-prognosis gynecologic disease with over 220,000 diagnoses each year worldwide and a 5-year survival rate less than 40%. In this study, we present a method for isolation and characterization of ovarian cancer cells from patient-derived ascites and tissue samples by using three-dimensional (3D) cell culture. Materials and Methods: Ascites and tissue samples were obtained from primary ovarian, peritoneal, and fallopian tube cancer patients intraoperatively. Samples were isolated and cultured within 6 hours after collection. After isolation, cells were resuspended in Matrigel and were placed at the center of each well. Optimized medium is added to each well. NanoCulture Plate LH96 (Low-Binding, MH pattern, 96-wells, Medical and Biological Laboratories Co., Ltd., Japan) were used for 3D cell culture. Plates were incubated at 37°C. The cultures were examined for metabolically active cells by ATP assay and medium changed on days 0, 1, 4, 7, 10, and 14. Results: We successfully obtained ascites-derived primary cell cultures within 1-7 days from 100% (3/31) of the ascites samples and 62% (5/8) from tissue-derived primary cell cultures. Spheroids-like structures were formed in 30% (1/3) of ascites samples and 50% (4/8) of tissue samples. The tumorigenicity and invasiveness of the cells were demonstrated using new 3D spheroid model cultured in vitro by NanoCulture Plate LH96. Conclusion: Primary ascites and tissue culture of ovarian cancer cells can successfully be cultured by 3D cell culture plates. We may apply this method for drug sensitivity testing; therefore, 3D cell culture has a potential to evaluate the sensitivity of candidate chemotherapeutic drug for individual patients. Citation Format: Yoshiko Nanki, Akira Hirasawa, Hiroyuki Nomura, Aki Okubo, Manabu Itoh, Tomoko Akahane, Tatsuyuki Chiyoda, Fumio Kataoka, Eiichiro Tominaga, Daisuke Aoki. Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A61.