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. 2022 Dec 8;207(8):1042–1054. doi: 10.1164/rccm.202207-1332OC

Figure 7.

Figure 7.

TanFe (transcellular activator of nuclear FOXF1 expression) increases angiogenesis in human vascular organoids (VOs) derived from a patient with ACDMPV. (A) Schematic diagram shows the differentiation protocol used to generate VOs from human induced pluripotent stem cells (hiPSCs). Brightfield images show morphological changes undergone by hiPSCs during VO differentiation. Scale bars for Days D0–D7, 500 μm. Scale bar for D15, 250 μm. (B) Images show immunostaining of whole-mount D15 VOs for CD31 (gray, endothelial cells) and PDGFRb (red, mural cells/pericytes). Nuclei were stained with DAPI. Scale bar, 200 μm. (C) TanFe treatment increases the percentages of CD31+ and PDGFRb+ cells in vascular organoids at D15 (n = 3 in each group). (D) Time course gene expression profile of FOXF1 during D3–D15 VO differentiation from human iPSCs. (E) Images show immunostaining of D15 VO sections for CD31 (gray) and FOXF1 (red). Scale bar, 200 μm. (F) High-magnification images of CD31-stained sections show lumen-like structures in D15 VOs (arrowheads). Scale bar, 50 μm. (G) Quantification of the endothelial density within VOs (n = 3). (H) Quantification of the number of lumen-like structures within VOs (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001. ACDMPV = alveolar capillary dysplasia with misalignment of pulmonary veins; BMP4 = bone morphogenetic protein 4; EB = embryoid body; FBS = fetal bovine serum; FGF2 = fibroblast growth factor 2; UD = undifferentiated human iPSCs; VEGF = vascular endothelial growth factor; YHWZ = tyrosine 3/tryptophan 5 -monooxygenase activation protein.