Fig. 3

Re-endothelialization of decellularized scaffold vasculature. a Method of human endothelial cell culture within the scaffold vasculature, using. 2 h of static culture followed by 3 days of arterial perfusion of culture prior to ex vivo function test or transplantation experiments. b Custom-built bioreactor allowing for endothelial cell deliveries to scaffold artery and vein, and subsequent perfusion culture. c Whole-mount fluorescence micrographs of a scaffold following 14 days of iPSC-derived epithelial cell culture and subsequent seeding with RFP-labeled HUVECs for an additional day. HUVECs distributed throughout the arterial and venous vascular systems (white arrowheads) with minimal leakage. Scale bar, 1 mm. d Vascular resistance of re-endothelialized intestinal scaffolds (n = 3) under constant rate perfusion. e Intestinal epithelium (CDX2⁺) and endothelium (CD31⁺) engrafted on an intestinal scaffold after 17 days culture in vitro. An open capillary vessel (white arrowhead) is seen between an artery and a vein. Scale bar, 100 µm. Error bars, mean ± s.d. of experimental values. *P < 0.05; **P < 0.01, Student’s t test. Ab, aboral end; O, oral end of intestine