The brain extracellular matrix (ECM) regulates myelin repair and regeneration after demyelination by interacting with neuronal progenitors and immune cells. Therefore, generating and characterizing ECM scaffolds in regions with distinct neuroregenerative capacities is crucial to identify factors that modulate cellular regenerative behavior. We established an effective decellularization protocol for the human neural stem cell (NSC)-rich subventricular zone (SVZ), frontal cortex (FC), and white matter (WM), and defined region-specific matrisomes using comparative proteomics. As proof of concept, we investigated the survival and differentiation of NSCs and monocytes within the decellularized scaffolds. NSCs cultured in WM and FC acquired an astrocytic phenotype, while both astrocytic and oligodendrocytic phenotypes were promoted by SVZ scaffolds. Additionally, monocytes seeded on SVZ and WM scaffolds exhibited an anti-inflammatory differentiation. This model is suitable for investigating ECM properties and assessing cell-matrix interactions.