Abstract 2159: Use of in vitro cell sheet engineering as a chemotherapeutic model for epithelial cancer

Abstract

Abstract Epithelial cancer is grown in the in vivo microenvironment mixed of keratinocytes, fibroblasts and matrix, vessels, and immune cells. The current in vitro 2-dimensional (2D) models for anti-cancer drug testing are commonly performed on a physical basis, resulting in significant difference from elucidating the real in vivo effects. To resolve the experimental gap, the 3D models have been developed to mimic the in vivo cancer microenvironment, while there is a lack of specific models for epithelial origin tumors. Therefore, we developed a new 3D model of using the in vitro cell sheet engineering for epithelial cancers and compared the results of several chemotherapeutic drug testing among in the 3D model, spheroid culture, and 2D-dish cell culture. The cell sheet included epithelial and subepithelial layers consisting of keratinocytes overlying the mixture of fibroblasts and endothelial progenitors. The spheroids of green fluorescent protein-tagged epithelial cancers were placed at the interface of epithelial and subepithelial layers. The spheroids with mixture of cancers and fibroblasts or cancer-associated fibroblasts (CAF) were made by the conventional hanging-drop method, and the 2D was simply seeded in the culture dish. The cell growth, viability, and hypoxia were calculated by cell counting kit (CCK), live/dead assay, propidium iodide and LOX-1 staining. The morphology, invasion, and mRNA and protein expressions were also compared among the different in vitro models. The results of live/dead assay and CCK showed higher viabilities in the cell sheet-based models before and after treatment with chemotherapeutic drugs. TGF-β and epithelial-mesenchymal transition-related mRNA and proteins were highly expressed in tumors growing in the cell sheet. Further, the incorporation of CAF into the cancer spheroids appeared to less decrease viable cancer cells by chemotherapeutic agents compared to the use of no or normal fibroblasts. Tumor angiogenesis and hypoxia were also apparently observed in the cell sheet model, which partly contributed to relative resistance to chemotherapeutic drugs. Taken together, the cell sheet-based cancer model might be applied to the in vitro observation of epithelial cancer growth and invasion, and anti-cancer drug testing. Citation Format: Eun Hye Kim, Jaewang Lee, Daiha Shin, Sunwoo Park, Minsu Kwon, Jong-Lyel Roh. Use of in vitro cell sheet engineering as a chemotherapeutic model for epithelial cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2159.