Abstract 3234: AlgiMatrix™ 3-D cell culture system as an in vitro tumor model for H460 non-small cell lung cancer cell line

Abstract

Abstract Objective: In cancer research, three dimensional (3D) cell culture models are rapidly gaining importance as they create a pragmatic microenvironment and mimic an in vivo system, which helps to understand cell-cell interactions. AlgiMatrix sponge (Invitrogen Corporation, Carlsbad, CA, USA) is a non toxic and biodegradable material made from alginate, which seems to construct a cell culture model that resembles normal cell characteristics and morphology. The main objective of the current study was to appraise the AlgiMatrix 3D cell culture system as a 3D invitro tumor model and to ascertain the anticancer activity of docetaxel. Methods: Cell titration was done to optimize seeding density (0.05, 0.10, 0.15, 0.25, 0.50, 0.75, 1.0, 1.25, 1.50, 3 & 6 million H460 cells/well using 6-well plate) and to determine the optimal incubation time for the formation of spheroids. On day 4, 9 & 13, the number and average size of the spheroids were determined using inverted phase contrast microscopy and imaging software. Further, optimized cell density (0.15 million cells/well) was used to determine the IC50 & IC90 values of docetaxel. The alamarBlue® assay was performed to determine the number of viable cells in control & treatment wells. Results: Optimized seeding density and the optimal incubation time was found to be 0.15 & 0.25 million H460 cells/well and 14 days respectively. Average size and the total number of the spheroids for 0.15 and 0.25 million H460 seeded cells was determined as shown in the table. Time PointsAverage size of spheroids (µm)Total Number of spheroids 0.15 million0.25 million0.15 million0.25 million4th day177.88 ± 61.79220.69 ± 61.2748 ± 477 ± 89th day205.35 ± 78.03232.90 ± 82.8790 ± 8189 ± 1513th day232.27 ± 89.07263.94 ± 86.40153 ± 18228 ± 17 Total number of cells obtained by alamarBlue® assay on 14th day was 38.64 ± 4.94 & 42.92 ± 3.29 million cells/well for 0.15 & 0.25 million seeded cells respectively. The IC50 & IC90 concentration of the docetaxel was found to be 193.19 µM & 290.47 µM when ascertained with 0.15 million cells/well. Conclusion: These results strongly support that the AlgiMatrix 3D Cell Culture System may be used as an in-vitro tumor model and to evaluate cytotoxicity of anticancer drugs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3234.