A 3D bioprinted neurovascular unit (NVU) model was developed to study glioblastoma (GBM) tumor growth in a brain-like microenvironment. The model includes human primary astrocytes, pericytes, and brain microvascular endothelial cells, with patient-derived GBM cells (JHH-520) used for the study. Fluorescence reporters and confocal high-content imaging were used to quantify real-time microvascular network formation and tumor growth. The model was validated using immunostaining, single-cell RNA sequencing (scRNAseq), and cytokine secretion analysis. scRNAseq revealed changes in gene expression and cytokine secretion associated with wound healing/angiogenesis, including the appearance of an endothelial mesenchymal transition (EndMT) cell population. The NVU-GBM model was used to test 18 chemotherapeutics and anti-cancer drugs to assess pharmacological relevance and robustness for high-throughput screening. The 3D tissue bioprinting (3DTBP) technology was used to assemble tissue biomimetics in a well with controlled spatial arrangements and desired cell type composition. 3DTBP is a promising tool for assembling complex tissue models due to its flexibility and reproducibility for large-scale production. It involves suspending cells in a polymeric bioink and using droplet- or extrusion-based techniques to create a customized xyz spatial pattern with selected cell types. 3DTBP has been successfully used for bioprinting various human tissues, including bone, cartilage, muscle, brain, liver, blood vessels, retina, and skin. It has also been shown to be a versatile tool for creating physiologically relevant disease tissue models, including modeling cancers. The NVU-GBM model was developed in a 96-well plate to create a patient-derived GBM growth assay in an NVU microenvironment amenable for high-throughput screening. The model was validated using immunostaining, fluorescence microscopy, and scRNAseq data. The model demonstrated high reproducibility and HTS compatibility, with a focused screen of 18 compounds tested in 4-point dose responses to assess anti-GBM potency and efficacy, as well as effects on the microvasculature network integrity. The NVU-GBM model was used to study GBM tumor growth and angiogenesis, with results showing increased outer-ring microvessels in the NVU-JHH520 co-culture group and decreased inward angiogenesis due to GBM presence. scRNAseq analysis revealed changes in gene expression and cytokine secretion associated with wound healing/angiogenesis, including the appearance of EndMT cells. The model also showed upregulation of inflammatory-related genes, maturation, and formation of junctions in endothelial cells and pericytes, myogenesis-related genes in pericytes, and epithelial mesenchymal transition in EndMTA 3D bioprinted neurovascular unit (NVU) model was developed to study glioblastoma (GBM) tumor growth in a brain-like microenvironment. The model includes human primary astrocytes, pericytes, and brain microvascular endothelial cells, with patient-derived GBM cells (JHH-520) used for the study. Fluorescence reporters and confocal high-content imaging were used to quantify real-time microvascular network formation and tumor growth. The model was validated using immunostaining, single-cell RNA sequencing (scRNAseq), and cytokine secretion analysis. scRNAseq revealed changes in gene expression and cytokine secretion associated with wound healing/angiogenesis, including the appearance of an endothelial mesenchymal transition (EndMT) cell population. The NVU-GBM model was used to test 18 chemotherapeutics and anti-cancer drugs to assess pharmacological relevance and robustness for high-throughput screening. The 3D tissue bioprinting (3DTBP) technology was used to assemble tissue biomimetics in a well with controlled spatial arrangements and desired cell type composition. 3DTBP is a promising tool for assembling complex tissue models due to its flexibility and reproducibility for large-scale production. It involves suspending cells in a polymeric bioink and using droplet- or extrusion-based techniques to create a customized xyz spatial pattern with selected cell types. 3DTBP has been successfully used for bioprinting various human tissues, including bone, cartilage, muscle, brain, liver, blood vessels, retina, and skin. It has also been shown to be a versatile tool for creating physiologically relevant disease tissue models, including modeling cancers. The NVU-GBM model was developed in a 96-well plate to create a patient-derived GBM growth assay in an NVU microenvironment amenable for high-throughput screening. The model was validated using immunostaining, fluorescence microscopy, and scRNAseq data. The model demonstrated high reproducibility and HTS compatibility, with a focused screen of 18 compounds tested in 4-point dose responses to assess anti-GBM potency and efficacy, as well as effects on the microvasculature network integrity. The NVU-GBM model was used to study GBM tumor growth and angiogenesis, with results showing increased outer-ring microvessels in the NVU-JHH520 co-culture group and decreased inward angiogenesis due to GBM presence. scRNAseq analysis revealed changes in gene expression and cytokine secretion associated with wound healing/angiogenesis, including the appearance of EndMT cells. The model also showed upregulation of inflammatory-related genes, maturation, and formation of junctions in endothelial cells and pericytes, myogenesis-related genes in pericytes, and epithelial mesenchymal transition in EndMT