Development and characterization of cancer stem cell-based tumoroids as an osteosarcoma model
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2020
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Abstract
Three-dimensional (3D) cancer tumor models are becoming vital approaches for high-throughput drug screening, drug targeting, development of novel theranostic systems, and personalized medicine. Yet, it is becoming more evident that the tumor progression and metastasis is fueled by a subpopulation of stem-like cells within the tumor that are also called cancer stem cells (CSCs). This study aimed to develop a tumoroid model using CSCs. For this purpose CD133+ cells were isolated from SaOS-2 osteosarcoma cell line with magnetic-activated cell sorting. To evaluate tumoroid formation ability, the cells were incubated in different cell numbers in agar gels produced by 3D Petri Dish® method. Subsequently, CD133+ cells and CD133− cells were co-cultured to investigate CD133+ cell localization in tumoroids. The characterization of tumoroids was performed using Live&Dead staining, immunohistochemistry, and quantitative polymerase chain reaction analysis. The results showed that, CD133+, CD133− and SaOS-2 cells were all able to form 3D tumoroids regardless of the initial cell number, but, while 72 hr were needed for CD133+ cells to self-assemble, 24 hr were enough for CD133− and SaOS-2 cells. CD133+ cells were located within tumoroids randomly with high cell viability. Finally, when compared to two-dimensional (2D) cultures, there were 5.88, 4.14, 6.95, and 1.68-fold higher messenger RNA expressions for Sox2, OCT3/4, Nanog, and Nestin, respectively, in CD133+ cells that were cultured within 3D tumoroids, showing longer maintenance of stem cell phenotype in 3D, that can allow more relevant screening and targeting efficiency in pharmaceutical testing. It was concluded that CSC-based tumoroids are propitious as 3D tumor models to fill the gap between conventional 2D in vitro culture and in vivo animal experiments for cancer research. © 2020 Wiley Periodicals LLC
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AC133 Antigen , Cell Line, Tumor , Humans , Models, Biological , Neoplastic Stem Cells , Osteosarcoma , Spheroids, Cellular , Tumor Cells, Cultured , Cell culture , Diseases , Polymerase chain reaction , RNA , Stem cells , Theranostics , Tumors , beta actin , CD133 antigen , nestin , octamer transcription factor 4 , transcription factor NANOG , CD133 antigen , PROM1 protein, human , Animal experiments , Immunohistochemistry , Osteosarcoma cells , Quantitative polymerase chain reaction , Stem cell phenotypes , Threedimensional (3-d) , Tumor progressions , Two Dimensional (2 D) , Article , cancer stem cell , cell selection , cell separation , cell viability , cellular distribution , controlled study , fluorescence microscopy , human , human cell , immunohistochemistry , in vitro study , osteosarcoma , osteosarcoma cell line , personalized medicine , phenotype , real time polymerase chain reaction , SaOS-2 cell line , tumor growth , tumor model , biological model , chemistry , cytology , metabolism , multicellular spheroid , osteosarcoma , tumor cell culture , tumor cell line , Diagnosis