Non‐neural extracellular matrix (ECM) has limited application in humanized physiological neural modeling due to insufficient brain‐specificity and safety concerns. Although brain‐derived ECM contains enriched neural components, certain essential components are partially lost during the decellularization process, necessitating augmentation. Here, it is demonstrated that the laminin‐augmented porcine brain‐decellularized ECM (P‐BdECM) is xenogeneic factor‐depleted as well as favorable for the regulation of human neurons, astrocytes, and microglia. P‐BdECM composition is comparable to human BdECM regarding brain‐specificity through the matrisome and gene ontology‐biological process analysis. As augmenting strategy, laminin 111 supplement promotes neural function by synergic effect with laminin 521 in P‐BdECM. Annexin A1(ANXA1) and Peroxiredoxin(PRDX) in P‐BdECM stabilized microglial and astrocytic behavior under normal while promoting active neuroinflammation in response to neuropathological factors. Further, supplementation of the brain‐specific molecule to non‐neural matrix also ameliorated glial cell inflammation as in P‐BdECM. In conclusion, P‐BdECM‐augmentation strategy can be used to recapitulate humanized pathophysiological cerebral environments for neurological study.