Figure 3.

In Vitro Angiogenesis Analysis of iPSC-Derived Endothelial Cells and Mural Cells

iPSC-derived ECs and MCs were cultured in Matrigel for in vitro assay of capillary tubule network formation.

(A) In vitro tubule formation by iPSC-ECs alone. CADASIL samples from two patients are in red and orange, respectively; control samples from two individuals are in blue and green, respectively.

(B) iPSC-ECs co-culture with iPSC-MCs. Result showing CADASIL iPSC-MCs failed to support stability of the tubule structure compared with the controls in the iPSC-EC/MC co-culture.

(C) In vitro tubule formation by iPSC-EC/MC co-culture in different combinations of CADASIL and control iPSC-ECs and iPSC-MCs. CADASIL iPSC-MCs had impaired ability to support the tubule stability formed by either CADASIL or control iPSC-ECs (warm colored lines) while the control iPSC-MCs were able to stabilize the tubule structure (cold colored lines).

(D) Immunofluorescence staining of the tubular structures formed by iPSC-EC/MC co-culture showing failure of CADASIL iPSC-MCs (α-SMA, green) to engage with iPSC-ECs (CD31, red) from 24 h onward during the angiogenesis assay.

Data in (A) to (C) are presented as mean ± SEM of three independent experiments (n = 3). Each experiment contained samples from three clones of each CADASIL or three clones of each control line. Two-way ANOVA with Tukey's post hoc test, ^∗∗∗p < 0.001. Scale bars, 50 μm.