Before endothelialization, an EB-based tissue featuring a SWIFT-printed single vessel was perfused with DMEM:F12 supplemented with 1% (v/v) of Matrigel for 20 min to enhance endothelial cell adhesion. HUVECs, cultured in EGM2 medium, were harvested from their flasks by incubating for 5 min in 0.05% trypsin/EDTA. The cells were centrifuged at 220g in DMEM with 10% FBS, strained, and resuspended at a density of 107 cells/ml in EGM2. The HUVEC suspension was perfused through the lumen at a flow rate of 40 μl/min until they could be seen exiting the tissue. The flow was then stopped, and the tissue gasket was laid flat for 10 min in the incubator to allow cells to settle at the surface of the lumen. This process was repeated three times, each time laying the gasket flat at a different angle such that the four quadrants of the lumen were coating with HUVECs. Tissues were then returned to being perfused with 1:1 mTSeR1:EGM2 medium at a flow rate of 40 μl/min for 24 hours before being fixed for analysis.

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