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. 2021 Sep 2;16(9):2159–2168. doi: 10.1016/j.stemcr.2021.08.003

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Quantitative assessment of the structural proprieties of hiPSC-VSMCs and primary mural cells in VoC

(A) Representative immunofluorescence images of microvascular network showing the hiPSC-ECs (magenta; agglutinin) and mural cells (green; SM22) derived vasculature unit spanning the complete length of microfluidic channel. Images showing hiPSC-ECs cultured with hiPSC-VSMCs, HBVSMCs, or HBVPs, respectively (10×). Scale bars, 200 μm.

(B) Representative confocal images of microvascular network showing hiPSC-ECs (gray; agglutinin) and mural cells (green; SM22). Images displaying xyz (i), xy (ii), and yz cross-sectional perspectives (iii). Images showing hiPSC-ECs cultured with hiPSC-VSMCs, HBVSMCs, or HBVPs, respectively (40×). Scale bars, 100 μm.

(C–E) Quantification of the percentage of mural cells associated with the hiPSC-EC lumen (% mural cells localized at the vascular network) (C), mean mural cell length (μm) (D), and mural cell circularity factor (the circle is 1) (E) in hiPSC-VSMCs, HBVPs, and HBVSMCs. Data are shown as ±SD from N = 3, n = 6; three independent experiments with two microfluidic channels per experiment.

(F) (Top) Representative confocal images of microvascular network showing hiPSC-ECs (gray; mCherry) and mural cells (red; SM22). (Bottom) Representative surface-rendered objects of confocal images showing microvascular network (gray; mCherry) and mural cells (colour-coded scale representing SM22 intensity). Images showing hiPSC-ECs cultured with hiPSC-VSMCs, HBVSMCs, or HBVPs, respectively (40×). Scale bars, 100 μm.

(G–I) Quantification of normalized mean cell SM22 intensity (G), number of SM22 + cells (H), and normalized mean cell SM22 intensity of mural cells in contact with hiPSC-ECs (I) in hiPSC-VSMCs, HBVPs and HBVSMCs. Intensity was normalized to hiPSC-VSMCs. Data are shown as ±SD from N = 3, n = 6; three independent experiments with two microfluidic channels per experiment.

One-way ANOVA (C–E and G–H) and two-way ANOVA (I) with Tukey's multiple comparison. ^∗p < 0.05, ^∗∗p < 0.001, ^∗∗∗p < 0.0001.

See also Figures S2E and S2F.