Glioblastoma multiforme (GBM) is recognized as the most common and lethal form of central nervous system cancer. GBM cells and astrocytes to high dosages of compounds with respect to efficacy and cytotoxicity, respectively, to discern the most efficacious drug for GBM. Among the 70 compounds tested, cediranib (a potent inhibitor of vascular endothelial growth factor (VEGF) receptor tyrosine kinases) exhibited the lowest cytotoxicity to astrocytes and high efficacy to GBM cells in a high-dose warmth map model. 1. Introduction Glioblastoma multiforme (GBM) is the most common, aggressive, and lethal main malignant brain tumor that stems from astrocytes. These tumors are usually highly malignant because the cells can metastasize from the primary tumor without detection and invade the surrounding normal brain tissue to form new tumor satellites that lead to tumor recurrence [1]. The current standard of care is surgical resection coupled with MLN2238 distributor ionizing radiation (IR) and the chemotherapeutic agent temozolomide (Temodar?, Temodal?, TMZ) [2, 3]. However, this treatment only provides patients with GBM a 12C14-month survival period after diagnosis [2, 3]. Despite aggressive surgical resection and chemotherapy, almost all patients with GBM present with tumor recurrence. Thus, many target-specific or general-chemotherapeutic brokers have been developed to remedy patients with GBM. Although some of the compounds exhibit good efficacy toward GBM, the producing cytotoxicity of normal glial cells in the central nervous system has been an issue. To measure cytotoxicity of compounds in normal glial cells, neural stem cells or astrocytes are used [4, 5]. Astrocytes are the most abundant member of the glial family and have a wide range of adaptive functions in the central nervous system (CNS). They interact with neurons, provide structural, metabolic, and trophic support, participate in synaptic activity, mediate ionic and transmitter homeostasis, and regulate blood flow [6, 7]. Since astrocytes play an important role in the CNS, treatment-induced toxicity of the CNS remains a major cause of morbidity in patients with malignancy [8]. Thus, a high-dose warmth map model comparing the responses of high-dose compounds on astrocytes and GBM cells is required to validate the most efficacious drugs toward GBM. Previous high-dose warmth map models using 2D cell-based high-throughput screening are well developed [9, 10]. However, because 2D cell-based assay does not fully reflect in vivo microenvironments (cell-to-cell and cell-to-extracellular matrix conversation), a 3D cell-based assay was used to screen compounds [11C15], including our previously developed system [13C17]. Especially, 3D cultured astrocyte and GBMs show more in vivo like model [18C20]. Thus, assay based on 3D cultured astrocyte and GBMs with high-throughput manner may give new potential to screen GBM target brokers. Our previous system [13C17] shows successfully data of 3D cell-based assays with high-throughput manner by comparing their own data with 2D cell-based MLN2238 distributor assay [13], gene [14], and clinical data [17]. By applying the abovementioned quantitative 3D-cultured cell-assay platform, astrocytes and patient-derived GBM cells were 3D-cultured and screened to select the most represented compounds that were not cytotoxic to normal brain cells and were particularly efficient for patient-derived GBM cells. Physique 1 shows 3D cell-based high-throughput screening chips culturing three-dimensionally four GBM cells and astrocyte. Since TMZ is usually a representative drug used in the treatment of patients with GBM, it was MLN2238 distributor used as a control compound to verify the high-dose warmth map. By comparing TMZ with 69 other compounds, compounds in the high-dose warmth map were tested for cytotoxicity and efficacy in GBM cells. Open in a separate window Physique 1 3D cell-based high-throughput screening chips. (a) Photo and schematic view of micropillar and microwell chip platform. Green dots are 3D-cultured astrocytes and glioblastoma multiforme (GBM) cells in alginate spot on the micropillar. (b) Schematic view of the experimental process. Cells are dispensed and immobilized in alginate onto the top of the micropillars and dipped MLN2238 distributor in the microwells made up of growth media for 1-day culture by sandwiching the micropillar and microwell chips. Compounds are dispensed into the microwells and cells Rabbit Polyclonal to FANCD2 are exposed to the compounds by moving the micropillar chip to a new microwell chip. 3D-cultured cells are stained with Calcein AM, and the.