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Anticancer drug discovery efforts have used 2‐D cell‐based assay models, which fail to forecast  in vivo  efficacy and result in a lower success rate of clinical approval. Recent 3‐D cell culture models are expected to bridge the gap between 2‐D and  in vivo  models. However, 3‐D cell culture methods that are available for practical anticancer drug screening have not yet been fully attained. In this study, we screened several polymers for their ability to suspend cells or cell spheroids homogeneously in a liquid medium without changing the viscosity behavior, and identified gellan gum ( FP 001), as the most potent polymer.  FP 001 promoted cell dispersion in the medium and improved the proliferation of a wide range of cancer cell lines under low attachment conditions by inhibiting the formation of large‐sized spheroids. In addition, cancer cells cultured with  FP 001‐containing medium were more susceptible to inhibitors of epidermal growth factor ( EGF ) signaling than those cultured under attachment conditions. We also showed that ligands of the  EGF  receptor family clearly enhance proliferation of  SKOV 3 ovarian carcinoma cells under anchorage‐independent conditions with  FP 001. Consistent with this result, the cells grown with  FP 001 showed higher  EGF  receptor content compared with cells cultured under attachment conditions. In conclusion, we developed a novel 3‐D cell culture system that is available for high throughput screening of anticancer agents, and is suitable for evaluation of molecular‐targeted anticancer drugs. Three‐dimensional cell culture using  FP 001 will be of value in the development of useful technologies for anticancer drug discovery.

Novel 3‐D cell culture system for in vitro evaluation of anticancer drugs under anchorage‐independent conditions