This study develops a 3D bioprinted neurovascular unit (NVU) model to investigate glioblastoma (GBM) tumor growth in a brain-like microenvironment. The NVU model includes human primary astrocytes, pericytes, brain microvascular endothelial cells, and patient-derived GBM cells (JHH-520). Fluorescence reporters and confocal high-content imaging are used to quantify real-time microvascular network formation and tumor growth. The model is validated through immunostaining for brain-relevant cellular markers and extracellular matrix components, single-cell RNA sequencing (scRNAseq) to establish transcriptomic changes, and secretion of NVU and GBM-relevant cytokines. ScRNAseq reveals changes in gene expression and cytokine secretion associated with wound healing and angiogenesis, including the appearance of an endothelial mesenchymal transition (EndMT) cell population. The NVU-GBM model is used to test 18 chemotherapeutics and anti-cancer drugs, assessing their pharmacological relevance and robustness for high-throughput screening (HTS). The bioprinted NVU-GBM model shows features of clinical GBM and is suitable for HTS in a 96-well multiwell plate format.This study develops a 3D bioprinted neurovascular unit (NVU) model to investigate glioblastoma (GBM) tumor growth in a brain-like microenvironment. The NVU model includes human primary astrocytes, pericytes, brain microvascular endothelial cells, and patient-derived GBM cells (JHH-520). Fluorescence reporters and confocal high-content imaging are used to quantify real-time microvascular network formation and tumor growth. The model is validated through immunostaining for brain-relevant cellular markers and extracellular matrix components, single-cell RNA sequencing (scRNAseq) to establish transcriptomic changes, and secretion of NVU and GBM-relevant cytokines. ScRNAseq reveals changes in gene expression and cytokine secretion associated with wound healing and angiogenesis, including the appearance of an endothelial mesenchymal transition (EndMT) cell population. The NVU-GBM model is used to test 18 chemotherapeutics and anti-cancer drugs, assessing their pharmacological relevance and robustness for high-throughput screening (HTS). The bioprinted NVU-GBM model shows features of clinical GBM and is suitable for HTS in a 96-well multiwell plate format.