Extended Data Fig. 3: Analysis of vascular organoids transplanted into mice.
From: Human blood vessel organoids as a model of diabetic vasculopathy
a, Intravenous infusion of the human-specific anti-CD31 (hCD31) antibody to label perfused human blood vessels (NC8) transplanted into immunodeficient NSG mice. Mouse vessels are visualized by a mouse-specific anti-CD31 antibody (mCD31). b, Functional human vasculature (detected by human-specific anti-CD31 immunostaining, red) in mice revealed by FITC–dextran perfusion (green). a, b, Experiments were repeated independently on n = 3 biological samples with similar results. c, Representative axial T2-weighted image, blood flow (perfusion), relative blood volume (rBV), mean transit time (MTT) and leakage (K2) measured by MRI. The axial plane was chosen so that both kidneys (outlined in white) and the implant (outlined in red) are visible. The analysed muscle tissue is outlined in green. Quantitative values (mean ± s.d.) for perfusion, relative blood volume, mean transit time and leakage are shown in the table. n = 3 mice. d, Representative low-magnification image of a transplanted vascular organoid stained for E-cadherin and human CD31. e, Arteriole (A) and venule (V) phenotypes appear within the human vascular organoid transplants (NC8). Representative haematoxylin- and eosin-stained histological sections are shown. f, Generation of human arterioles (A) shown by staining for human-specific CD31 (endothelial cells), tightly covered with vascular smooth muscle cells detected by SMA, calponin 1 and human-specific MYH11 immunostaining. As a control, endothelial cells of mouse kidney arterioles do not crossreact with the human-specific CD31 antibody (top right). Samples were also stained with DAPI. g, h, Arterioles of human origin (hCD31+) express arterial markers ephrinB2 and DLL4. i, j, Human venous structures (hCD31+) express the venous markers EPHB4 and COUPTF2. k, BFP-tagged vascular organoids (H9) were transplanted into mice. Staining of the organoids with a human-specific CD31 (hCD31) antibody reveal the human identity of the cells. A representative image from n = 5 mice is shown. d–k, Experiments were repeated independently on n = 3 biological samples with similar results. DAPI was used to counterstain nuclei. Scale bars, 20 μm (e–h), 50 μm (i, j), 100 μm (b, k), 200 μm (a) and 500 μm (d).
