Fig. 3

Citrated whole-blood perfusion in engineered microvessels (µVs). a Brightfield stitched large image of red blood cell-filled pattern and sprouts with magnified view (inset, white dotted boundary) for human embryonic stem cell-derived endothelial cell (hESC-EC)-seeded µV only constructs after 4 days of culture. Scale bar, 200 μm. b Maximum intensity projection of confocal z-stack of constructs with adhered platelets after 30-min perfusion and subsequent phosphate-buffered saline (PBS) washes for untreated hESC-EC-seeded constructs (top) and phorbol myristate acetate (PMA)-treated hESC-EC-seeded constructs (bottom) stained for CD31 (red) and CD41a (green). Scale bar, 200 μm. c Quantification of platelet adhesion on the vessel wall for constructs seeded with human umbilical vein endothelial cells (HUVECs) in control conditions (C–HUVEC), hESC-ECs in control conditions (C–hESC-EC), and hESC-ECs treated with PMA (PMA–hESC-EC) or interleukin (IL)-1β (IL-1β–hESC-EC). Data are expressed as a percentage of the vessel wall surface area. N = 2, 2, 2, and 3 biologically independent samples for C–HUVEC, C–hESC-EC, PMA, and IL-1β, respectively. Representative images for a, b from two biologically independent samples of C–hESC-ECs and two biologically independent samples of PMA–hESC-EC, with similar results. Error bars, mean ± SEM