Abstract
In evaluation of cell viability and apoptosis, spatial heterogeneity is quantified for cancerous cells cultured in 3-D in vitro cell-based assays under the impact of anti-cancer agents. In 48-h experiments using human colorectal cancer cell lines of HCT-116, SW-620, and SW-480, incubated cells are divided into control and drug administered groups, to be grown in matrigel and FOLFOX solution, respectively. Our 3-D cell tracking and data acquisition system guiding an inverted microscope with a digital camera is utilized to capture bright field and fluorescent images of colorectal cancer cells at multiple time points. Identifying the locations of live and dead cells in captured images, spatial point process and Voronoi tessellation methods are applied to extract morphological features of in vitro cell-based assays. For the former method, spatial heterogeneity is quantified with the second-order functions of Poisson point process, whereas the deviation in the area of Voronoi polygons is computed for the latter. With both techniques, the results indicate that the spatial heterogeneity of live cell locations increases as the viability of in in vitro cell cultures decreases. On the other hand, a decrease is observed for the heterogeneity of dead cell locations with the decrease in cell viability. This relationship between morphological features of in vitro cell-based assays and cell viability can be used for drug efficacy measurements and utilized as a biomarker for 3-D in vitro microenvironment assays.
Highlights
C ELL line and tissue based in-vitro cell culture systems are tools to emulate in-vivo cell behavior and cellular interactions [1]
To study the relationship between cell viability and spatial heterogeneity in the positions of cancer cells, we present three case studies using colorectal cancer cell lines of HCT116, SW-620 and SW480
Control group of colorectal cancer cells are cultured in metragel, whereas the cells of the treatment group are in FOLFOX solution, and both are provided with nutrition and gas supplies at an ideal temperature of 37◦C for 48 hours
Summary
C ELL line and tissue based in-vitro cell culture systems are tools to emulate in-vivo cell behavior and cellular interactions [1]. Viability of incubated cells under the impact of anti-cancer drugs and their morphology changes can be observed via digitized microscopic images from cell cultures captured during in-vitro experiments. Ripley‘s K-function and its derived versions can be used to test the consistency of observed patterns with a homogeneous Poisson process [7] Voronoi tessellation is another spatial analysis tool for partitioning an Euclidian space into subregions based on node locations, where an association of subregions of a given plane to the closest nodes results in a tessellation diagram containing information specific to a specific plane [8]. We examine the relationship between cell viability and morphological features of 3D microenvironment using spatial analysis methods, namely poisson point process and Voronoi tessellations.
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