Transplantation of hPSC‐Derived Pericyte‐Enriched Vascular Assemblies Restores Cerebral Perfusion in the Context of Permanent Brain Ischemia

ABSTRACT Despite advancements in vascular reconstruction techniques, insufficient distal perfusion following ischemic stroke continues to cause irreversible neurological damage, and effective strategies for reconstructing functional microvascular networks remain inadequate. While vascular organoids have shown potential, their clinical translation is limited by the need for invasive surgical resection to accommodate large 3D grafts. This study improves the vascular organoid technology pipeline by developing hPSC‐derived Pericyte‐Enriched Vascular Assemblies. These assemblies are composed of CD31 + endothelial cells, PDGFRβ + pericytes, and SMA‐α + smooth muscle cells, with pericytes accounting for 62.6% of the composition, making them the key cells determining the vascular organoid phenotype. After transplanting single‐cell suspensions derived from these assemblies into the infarcted area, cerebral blood flow perfusion in mice was significantly restored, and sensory function improved. Mechanistic studies revealed that ischemic stress specifically upregulated the PRKCA signaling pathway in pericytes. Knockout of this kinase impaired the phenotype of the grafts, confirming its role as a master regulator of pericyte‐mediated vascular remodeling. This study shifts the paradigm from structural biomimicry to function‐driven design, highlighting the central regulatory role of pericytes in vascular organoid repair and providing new perspectives for cellular therapy and clinical treatment of ischemic stroke.