The endothelial functionality of the resulting BLECs was assessed via tube formation assays.<h4>Results</h4>Immunocytochemical analysis confirmed the expression of key BMEC markers, including ZO-1, CD31, and occludin, showing both phenotypic and functional characteristics of brain microvascular endothelium.<h4>Conclusion</h4>This MSC-based differentiation approach provides a robust and physiologically relevant in vitro BBB model with potential applications in studying neurological disease mechanisms and screening therapeutic agents. Here, PECAM1 is linked to nervous system disorder.