Effects of seeding density and collagen stiffness on nuclei clustering pattern of diffuse type of stomach cancer

Abstract

Gastric cancer also known as stomach cancer is the second and fourth most common cause of can-cer death in males and females respectively throughout the world. Especially, mechanism underlying diffuse type gastric cancer (DGC) formation is not fully understood. Therefore, understanding behavior of the for-mation of cluster as aggregation of HS746T, one of DGC cell line in vitro could contribute to develop therapeu-tic strategies of DGC. In this study, we tried to identify the spatial distribution in terms of cluster, which implies cell-cell sig-naling. Cell signaling can be classified by the distance between cells such as autocrine signals themselves, jux-tacrine signals between adjacent or touching cells through gap junctions and paracrine signals in the vicinity. In order to figure out cluster to understand visualized cell signaling, we analyzed images using mean-shift clustering method at ‘Fiji’ software. We standardized the methods to analyze nuclei clustering pattern by increasing seeding density and stiffness, which can presume the progress of tumor cell growth. We confirmed the cluster area at high cell confluency was relatively bigger than low cell confluency image. Interestingly, even low cell confluency had relatively high cluster area because biased and compact cell distribution enables to diffuse signaling molecules effectively. In conclusion, cluster analysis just by the images is expected to substitute the additional proliferation assay to understand interactions between cell network. Thus, quantified cluster area and number caused by cell signaling may assist future studies to presume and identify tumor aggregation or metastasis.