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Bio Protoc. 2018 Apr 20;8(8). pii: e2815. doi: 10.21769/BioProtoc.2815.

3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells.

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Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Rd II, Guangzhou 510080, China.
Department of Pathology and the Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT06520, USA.


Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated spheroid and clinical pathology of ovarian cancer, and that TAMs promote spheroid formation and tumor growth at early stages of transcoelomic metastasis in orthotopic mouse models. We have established an in vitro spheroid formation assay using a 3D co-culture system in which mouse GFP+F4/80+CD206+ TAMs isolated from spheroids of ovarian cancer-bearing donor tomatolysM-cre mice were mixed with ID8 cells (TAM:ID8 at a ratio of 1:10) in medium containing 2% Matrigel and seeded onto the 24-well plate precoated with Matrigel. As transcoelomic metastasis is also associated with many other cancers such as pancreatic and colon cancers, TAM-mediated spheroid formation assay would provide a useful approach to define the molecular mechanism and therapeutic targets for ovarian cancer and other transcoelomic metastasis cancers.


3D co-culture system; Ovarian cancer; Spheroid formation; Transcoelomic metastasis; Tumor-associated macrophage

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