Fabrication of 3D calcium‐alginate scaffolds for human glioblastoma modeling and anticancer drug response evaluation

Abstract

The three-dimensional (3D) cell culture model has been increasingly used to study cancer biology and screen for anticancer agents due to its close mimicry to in vivo tumor biopsies. In this study, 3D calcium(Ca)-alginate scaffolds were developed for human glioblastoma cell culture and an investigation of the responses to two anticancer agents, doxorubicin and cordycepin. Compared to the 2D monolayer culture, glioblastoma cells cultured on these 3D Ca-alginate scaffolds showed reduced cell proliferation, increased tumor spheroid formation, enhanced expression of cancer stem cell genes (CD133, SOX2, Nestin, and Musashi-1), and improved expression of differentiation potential-associated genes (GFAP and β-tubulin III).Additionally, the vascularization potential of the 3D glioblastoma cells was increased, as indicated by a higher expression of tumor angiogenesis biomarker (VEGF) than in the cells in 2D culture. To highlight the application of Ca-alginate scaffolds, the 3D glioblastomas were treated with anticancer agents, including doxorubicin and cordycepin. The results demonstrated that the 3D glioblastomas presented a greater resistance to the tested anticancer agents than that of the cells in 2D culture. In summary, the 3D Ca-alginate scaffolds for glioblastoma cells that were developed in this study offer a promising platform for anticancer agent screening and the discovery of drug-resistant mechanisms of cancer. K E Y W O R D S3D brain cancer, 3D calcium-alginate scaffolds, anticancer drug screening, brain cancer model, glioblastoma

Transcriptomic Profiling of 3D Glioblastoma Tumoroids for the Identification of Mechanisms Involved in Anticancer Drug Resistance

“…The limitations of 2D cell culture might be overcome by culturing cells under more physiological 3D conditions. Thus, 3D in vitro tumor models would be powerful and effective tool, in cancer research and low-cost anticancer drug screening (10,11). Closely imitating the complex in vivo tumor microenvironment helps researchers to reveal and understand the underlying molecular mechanism for tumorigenesis, development, proliferation, migration, invasion, angiogenesis, and resistance to anticancer drugs.…”

“…Fabrication of porous scaffolds. Ca-alginate scaffolds for 3D cell culture were fabricated using freeze-drying technique as described in our previous study (11). Briefly, 2% sodium alginate was dissolved in sterile deionized (DI) water.…”

“…In order to recapitulate such cell microenvironments in vivo, three-dimensional (3D) culture has been alternatively used as in vitro tumor models, which can simulate in vivo cell behaviors, and provide more comparable and reliable results (8). Several 3D in vitro glioblastoma models have been established using extracellular matrix-based scaffolds and elucidated the dramatic differences in terms of cell proliferation, morphology, and drug resistance between 2D and 3D glioblastoma cells in culture (9)(10)(11). Our previous study reported the up-regulation of genes associated with stemness and differentiation, and vascular endothelial growth factor (VEGF) angiogenesis factor in 3D glioblastoma, which might play important roles in the enhancement of drug resistance (11).…”

“…Several 3D in vitro glioblastoma models have been established using extracellular matrix-based scaffolds and elucidated the dramatic differences in terms of cell proliferation, morphology, and drug resistance between 2D and 3D glioblastoma cells in culture (9)(10)(11). Our previous study reported the up-regulation of genes associated with stemness and differentiation, and vascular endothelial growth factor (VEGF) angiogenesis factor in 3D glioblastoma, which might play important roles in the enhancement of drug resistance (11). Recently, several investigators have examined the effects of glioblastoma multiforme tumor cells culture condition on gene expression profiles.…”

Engineering Three-Dimensional Tumor Models to Study Glioma Cancer Stem Cells and Tumor Microenvironment

“…Furthermore, 3D cultures also showed greater resistance to alkylating agents with a high regulation of MGMT (Lv et al, 2016 ). Different scaffolds created with other relevant tumor ECM components such as HA as well as with synthetic materials have also been described (Erickson et al, 2018 ; Chaicharoenaudomrung et al, 2019 ; Xiao W. et al, 2019 ).…”