Volumetric Analysis of 3-D-Cultured Colonies in Wet Alginate Spots Using 384-Pillar Plate. (June 2018)

• Record Type: Journal Article
• Authors: Lee, Dong Woo; Choi, Yea-Jun; Lee, Sang-Yun; Kim, Myoung-Hee; Doh, Il; Ryu, Gyu Ha; Choi, Soo-Mi
• Other Names: Jeyasekharan Anand D. guest-editor.
• Is Part Of: SLAS technology. Volume 23:Number 3(2018)
• Page Start: 226
• Page End: 230
• Publication Date: 2018-06
• Subjects: 3-D cell culture -- cell encapsulation with alginate -- high-throughput screening -- assay miniaturization -- clonogenic assay; Medical laboratory technology -- Periodicals; Laboratories -- Equipment and supplies -- Periodicals; Diagnosis, Laboratory -- Periodicals; 616.075
• DOI: 10.1177/2472630317737698
• Languages: English
• ISSNs: 2472-6303

Abstract

The volumetric analysis of three-dimensional (3-D)-cultured colonies in alginate spots has been proposed to increase drug efficacy. In a previously developed pillar/well chip platform, colonies within spots are usually stained and dried for analysis of cell viability using two-dimensional (2-D) fluorescent images. Since the number of viable cells in colonies is directly related to colony volume, we proposed the 3-D analysis of colonies for high-accuracy cell viability calculation. The spots were immersed in buffer, and the 3-D volume of each colony was calculated from the 2-D stacking fluorescent images of the spot with different focal positions. In the experiments with human gastric carcinoma cells and anticancer drugs, we compared cell viability values calculated using the 2-D area and 3-D volume of colonies in the wet and dried alginate spots, respectively. The IC50 value calculated using the 3-D volume of the colonies (9.5 μM) was less than that calculated in the 2-D area analysis (121.5 μM). We observed that the colony showed a more sensitive drug response regarding volume calculated from the 3-D image reconstructed using several confocal images than regarding colony area calculated in the 2-D analysis.