Figure 2.

Mechanism of the intra-microvascular network formation

(A) Schematic of the experimental setup. MTs were generated combining hiPSC-CMs, hiPSC-ECs (green, GFP), and hiPSC-CFs. These MTs were cocultured with hiPSC-ECs (red, mCherry) and HBVPs in chips.

(B and C) Representative images of outside-in (B) and inside-out (C) anastomosis and formation of hybrid vessels by interconnection of internal microvascular network (green, GFP) and external vascular network (red, mCherry) (20×). Scale bar, 150 μm.

(D and E) Representative confocal images of hybrid vessels visible in (B) and (C), respectively. Internal hiPSC-ECs (green, GFP) and external hiPSC-ECs (orange, mCherry). Images displaying maximum projection in xyz (i), xy (ii), and yz cross-sectional perspectives (iii) (40×). Scale bar, 100 μm. Arrows indicate the anastomosed points and hybrid lumens.

(F) Quantification of number of vascularized MTs (%, number vascularized MTs/total number of MTs). MTs contained CMs from three different hiPSC lines. Error bars are shown as mean ± SD from N = 3; three independent experiments with at least 10 MTs in each experiment. One-way ANOVA; ns, not significant.

(G) Quantification of number of anastomosed MTs (%, #anastomosed MTs/total # of MTs) in independent experiments. Error bars are shown as mean ± SD from N = 6; six independent experiments with at least nine MTs in each experiment. Student’s t test, ∗p < 0.05.

See also Figure S2.