The perigraft compartment is the dominant cellular source for decellularized tissue-engineered vascular graft...

Compared with conventional synthetic grafts, decellularized tissue-engineered vascular grafts (dTEVGs) hold transformative potential for hemodialysis access owing to their capacity for host‑driven recellularization. To investigate the cellular origin of this process, we established a rat arteriovenous graft model using dTEVGs and systematically mapped the temporal dynamics of infiltrating cell phenotypes. To spatially resolve cellular origins, we developed a specifically designed strategy featuring surface-specific cytophobic barriers that selectively block cellular entry from defined compartments. Our results demonstrated that abluminal blockade reduced cellular infiltration by 96.7 % and abolished collagen deposition, whereas luminal blockade showed no significant effect, definitively establishing the perigraft compartment as the dominant cellular source for dTEVG recellularization. Crucially, perigraft‑sourced cells exhibited superior proliferative capacity, stemness, and pro‑regenerative potential, making them uniquely suited for driving functional vascular regeneration. These findings, enabled by a compartment-selective blockade model, definitively establish the perigraft compartment as the dominant cellular source for recellularization in this dTEVG and underscore the critical importance of perivascular niche engagement. This work provides a methodological framework and a theoretical rationale for considering abluminal-focused strategies in the design of next-generation dTEVGs. STATEMENT OF SIGNIFICANCE: This study definitively establishes the perigraft compartment, not the lumen, as the dominant cellular source for recellularizing decellularized tissue-engineered vascular grafts. These perigraft-derived cells exhibit superior regenerative potential, overturning the long-held circulation-centric paradigm and guiding the design of next-generation grafts for improved hemodialysis access.