Fig 2: Human iPSC-EC-derived microvascular meshes enhance vascularization. a Fluorescent images of iPSC-EC meshes with different geometries (square, pentagon, hexagon, and octagon). b iPSC-EC meshes with more complex patterns (spider web and capillary bed). c Confocal images of an iPSC-EC mesh at the contracted and junction regions. iPSC-EC expressing GFP is green, human CD31 antibody is red, and nucleus is blue. All samples were imaged after 2 days of culture. d Hematoxylin/eosin and immunostaining images of retrieved devices after 2 weeks of subcutaneous implantation in SCID-Beige mice. In all in vivo experiments, NHDFs were mixed with iPSC-ECs (iPSC-EC:NHDF = 9:1). The yellow arrowheads point to blood vessels with erythrocytes inside. The white dash lines mark the interface between the device and panniculus carnosus muscle. Mouse CD31 antibody is green and a-smooth muscle actin (a-SMA) is red. e The density and area percentage of blood vessels at the interface. n = 5 in all groups. Data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and NS (P > 0.05) no significant difference. One-way analysis of variance. f Immunostaining images of human (red) and mouse CD31 (green) antibodies showing the anastomoses between iPSC-EC derived vessels and mouse vessels

Fig 3: Improvement of re-vascularization of rat islets and diabetes correction in SCID-Beige mice. a Schematics and a microscopic image of rat islets (green) in a Mesh device. Microvascular mesh is red and nylon grid is blue. b Non-fasting blood glucose (BG) concentration of the mice after transplantation. Grafts were retrieved after different time points. Most of the mice were kept alive for 1 week after retrieval while 3 mice from the Mesh group were used for perfusion studies prior to retrievals. During 42 days of transplantation (n = 9 in No cell, n = 11 in Random, and n = 14 in Mesh): *P < 0.05 and NS (P > 0.05) no significant difference. ANCOVA, time was treated as continuous covariate. c BG concentrations during intraperitoneal glucose tolerance tests (IPGTT) after 30 and 90 days of transplantation. IPGTT at Day 30 (n = 6 in Normal mice and No cell, n = 8 in Random and Mesh). Data are mean ± SEM. *P < 0.05 and NS (P > 0.05) no significant difference. ANCOVA, time was treated as continuous covariate. d Hematoxylin/eosin staining of rat islets and blood vessels in retrieved devices (Day 42) and the number of blood vessels around a rat islet (within a 200 µm distance). Yellow arrowheads point to blood vessels with erythrocytes inside. White arrows point to rat islets. The round material in the cross-sections is nylon grid. No cell group consists of 11 islets pooled from 3 mice; Random group consists of 13 islets pooled from 3 mice; Mesh group consists of 12 islets pooled from 3 mice. *P < 0.05, ***P < 0.001, and NS (P > 0.05) no significant difference. One-way analysis of variance. e Cross-sectional immunostaining images of rat insulin (red) and mouse blood vessels (CD31, green) in a retrieved Mesh device. f Immunostaining images (parallel section) using human (red) and mouse (green) CD31 antibodies indicate the anastomoses between human and mouse blood vessels. g Confocal image of perfused blood vessels in re-vascularized rat islets in the Mesh group after 42 days of transplantation. h Fluorescent images of re-vascularized rat islets after 91 and 112 days of transplantation from the Mesh group. The rat islets expressing GFP are green and perfused blood vessels are red

Fig 4: Inhibition of xenograft tumor growth and metastasis by matrine in vivo.a Time course of tumor growth, measured as tumor volume in each group at the indicated time of treatment with normal saline or matrine (100 mg/kg, ip). Data are presented as mean ± SD (n = 3, *P < 0.05). b Tumors in matrine-treated mice were significantly smaller than those in normal saline-treated mice. c The bar graph represented the mean of tumor weight from matrine-treated and control mice (n = 3, *P < 0.05). d The representative histological examinations of the dissected tumors with matrine or normal saline treatment using H&E staining, TUNEL assay, Ki-67 and CD31 antibodies. Scale bar: 50 µm. e The global phosphorylation levels of tyrosine and serine/threonine were significantly decreased in matrine-treated A2780 cells and in matrine-treated tumor tissue lysates, respectively. f, g Western blot and IHC staining demonstrated a decrease in the phosphorylation levels of ERK1/2, MEK1/2, PI3K, Akt, mTOR, FAK, RhoA, VEGFR2, and Tie2 in xenografts treated with matrine compared with those in control group. Scale bar: 50 µm. h A2780 cells were injected into tail veins of BALB/c nude mice. Representative images showed lungs with metastatic nodules from the matrine-treated mice and the control group. The left panel presents macroscopic appearances of lung metastatic nodules, and the middle and right panels present the H&E staining of lung tissues in mice with matrine or normal saline treatment. i The number of lung metastatic nodules was significantly reduced in mice treated with matrine compared with the control group. j H&E staining of hepatic tissues in mice with matrine or normal saline treatment. Scale bar: 50 µm. k Concentrations of ALT and AST were measured in the blood at the end of matrine treatment. Data are presented as mean ± SD

Fig 5: Improved diabetes correction by the iPSC-EC microvascular meshes in SCID-Beige mice. a A microscopic image of rat islets in an iPSC-EC mesh device. The gray color aggregates are rat islets. The white arrow points to iPSC-EC mesh. In all in vivo experiments, NHDFs were mixed with iPSC-ECs (iPSC-EC:NHDF = 9:1). b Non-fasting BG concentration of the mice during 91 days of subcutaneous transplantation (n = 5 in No iPSC-EC, n = 6 in Random iPSC-EC and Mesh iPSC-EC). *P < 0.05. ANCOVA, time was treated as continuous covariate. c BG concentration during intraperitoneal glucose tolerance test (IPGTT) after 30 and 90 days of transplantation. IPGTT at Day 30 (n = 6 in Normal mice, Random iPSC-EC, and Mesh iPSC-EC, and n = 4 in No iPSC-EC). Data are mean ± SEM. *P < 0.05. NS (P > 0.05) no significant difference. IPGTT at Day 90 (n = 6 in Normal mice, n = 3 in No iPSC-EC, n = 4 in Random iPSC-EC, and n = 6 in Mesh iPSC-EC): *P < 0.05. ANCOVA, time was treated as continuous covariate. d Immunostaining images (parallel section) of human (red) and mouse CD31 (green) antibodies indicating the anastomoses between human and mouse blood vessels. e Confocal image of perfused blood vessels in a re-vascularized rat islet retrieved from the Mesh group at Day 91. f Hematoxylin/eosin staining image of a retrieved iPSC-EC mesh device at Day 91. Black arrow points to a rat islet and yellow arrowheads point to blood vessels containing erythrocytes. g Immunostaining image of insulin and blood vessels surrounding rat islets in a retrieved iPSC-EC mesh device at Day 91. Rat insulin antibody is red and mouse CD31 antibody is green