endothelial marker cd31 pecam 1  (R&D Systems)

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: High-throughput (HT)-compatible and nearly xeno-free synthesis of vascular networks and blood vessel organoids from fluorescently tagged human induced pluripotent stem cells (hiPSCs). A Schematic illustration of the new differentiation protocol and representative images for the main differentiation steps (scale bars: d-2, d0, d3, d5 = 100 µm; d7, d12 = 250 µm; d14, d17 = 500 µm). Created in BioRender. Skowronek, D. (2025) https://BioRender.com/e70e302 . B Shown are the steps of embedding the vascular aggregates in an Akura 96-well plate and transferring the vascular networks from the Akura 96-well plate to a PrimeSurface 96 Slit-well plate. C The use of PrimeSurface 96 Slit-well plates reduces the time required for medium exchange (left image). Akura 96-well plates allow aggregates to be embedded in small cavities, minimizing the matrix surrounding the vascular networks (black arrows) and allowing direct transfer of vascular networks (white arrows) to new plates without time-consuming manual extraction of the networks from the gel (middle and right images). D The new protocol is simple to handle and achieves high synthesis efficiency after minimal training. Shown are the efficiencies of three training runs. E The sprouting efficiency is maintained when fetal bovine serum (FBS) is replaced with human platelet lysate (hPL) or chemically defined Panexin CD (PCD). The total numbers of sufficiently sprouted networks and vascular aggregates with insufficient sprouting are written inside the bars. F,G HiPSC-derived vascular networks (F) and blood vessel organoids (G) differentiated with the HT-compatible and nearly xeno-free protocol consist of a complex network of endothelial cells (CD31) and associated pericytes (PDGFR-β) [representative images; scale bars: 50 µm ( F ); 200 µm ( G )]. White arrowheads indicate angiogenic sprouts. H Perfusion of vascular networks with TMR-amino-dextran in OrganoPlate graft plates shows anastomoses between the GFP-labeled vascular networks and the HUVEC-derived vascular bed (top, white arrowheads) as well as correct formation and permeability of the vascular networks (bottom, scale bar: 50 µm)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: High Throughput Screening Assay, Transferring, Extraction, Derivative Assay, Labeling, Permeability

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: Perfusion of blood vessel organoids (BVO) on chorioallantoic membranes (CAM). A Schematic illustration of the perfusion approach. Created in BioRender. Skowronek, D. (2025) https://BioRender.com/n06n764 . B Shown are blood vessel organoids cultivated on the CAM associated with chicken blood vessels. The pictures were taken on day 1 and day 6 (scale bars = 2 mm). C Sectioning and H&E staining demonstrated nucleated chicken erythrocytes within the vascular structures of the blood vessel organoid (black arrow head) (upper scale bar = 500 µm; bottom scale bar = 25 µm). D The expression of CD31 (upper image) and PDGFR-β (lower image) were verified by immunohistochemistry staining (brown) (scale bar = 100 µm)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Angiogenesis

Article Title: High-throughput differentiation of human blood vessel organoids reveals overlapping and distinct functions of the cerebral cavernous malformation proteins

doi: 10.1007/s10456-025-09985-5

Figure Lengend Snippet: Structural defects in KO vascular networks. A Immunofluorescence staining for CD31 (endothelial marker, green) and PDGFR-β (pericyte marker, red) indicated a reduced correlation between ECs and pericytes in CCM1, CCM2, and CCM3 KO vascular networks (scale bar: 100 µm). Correlation was evaluated by using the Pearson’s correlation coefficient (r) calculated with the JACoP ImageJ plugin. B , C Immunofluorescence staining for ZO-1 (B) and VE-cadherin ( C ) revealed irregular tight and adherens junctions in CCM1 , CCM2, and CCM3 KO vascular networks (Alexa 647; scale bars: 25 µm). Statistical analyses demonstrated a significant reduction of Alexa 647 (ZO-1) fluorescence intensity in CCM2 and CCM3 KO networks. Data are presented as individual data points and means. Multiple two-sample t-tests with Welch's correction and Holm-Šídák adjustment for multiple testing were used for statistical analyses (* = Padj < 0.05)

Article Snippet: To confirm endothelial differentiation, cells were fixed with 4% PFA at passage 1 and stained for the endothelial markers CD31 (Cell Signaling, 3528S, 1:800), VE-cadherin (2500S, 1:400, Cell Signaling) and VWF (Thermo Fisher Scientific, MA5-14029, 1:66).

Techniques: Immunofluorescence, Staining, Marker, Fluorescence

Journal: Alzheimer's & dementia : the journal of the Alzheimer's Association

Article Title: tPA supplementation preserves neurovascular and cognitive function in Tg2576 mice.

doi: 10.1002/alz.13878

Figure Lengend Snippet: FIGURE 4 Intranasal tPA administration attenuates pial and neocortical CAA in aged Tg2576 mice. A, In vivo two-photon excited fluorescent images illustrating methoxy-XO4+ amyloid deposits (magenta) around somatosensory cortex blood vessels (green) identified by retroorbital injection of 70 kDa Texas Red dextran. Methoxy-XO4+ Aβ deposits are reduced in intranasal tPA-treated Tg2576 compared to vehicle-treated Tg2576 mice. B, CAA burden (% thioflavin-S+ area) in cortical sections labeled with thioflavin-S and the blood vessel marker CD31 is reduced in intranasal tPA-treated Tg2576 compared to vehicle-treated Tg2576 mice, but amyloid plaque burden (% thioflavin-S+ plaque area) was comparable between the two groups. In (A) and (B), arrows indicate CAA and arrowheads plaques. N = 5/group; unpaired t test; scale bars, 100 μm in (A) and 200 μm in (B); data presented as mean ± standard error of the mean. Aβ, amyloid beta; CAA, cerebral amyloid angiopathy; tPA, tissue plasminogen activator; WT, wild type.

Article Snippet: For hepcidin immunolabel- ing, coronal sections were co-stained with hepcidin (rabbit polyclonal, 1:200, Catalog# ab30760, abcam) and the endothelial marker CD31 (goat polyclonal, 1:150, Catalog # AF3628, R&D Systems) followed by thioflavin-S (0.5%), mounted on slides, and imaged with a confocal microscope (Leica SP8).

Techniques: In Vivo, Injection, Labeling, Marker

Journal: Biomedicines

Article Title: External Support of Autologous Internal Jugular Vein Grafts with FRAME Mesh in a Porcine Carotid Artery Model

doi: 10.3390/biomedicines12061335

Figure Lengend Snippet: Microscopical examinations of autologous internal jugular vein interposition grafts in porcine carotid arteries at one month post implantation (control group). Representative cross-sections of the midgraft regions are shown in Pigs 2 ( A – D and U – X ), 3 ( E – H ), 4 ( M – P ), 5 ( Y – Z2 ), 6 ( I – L ), and 7 ( Q – T ). The seven control samples are ordered from left to right according to degree of pathological remodeling and vein graft lesions, i.e., patent, mildly stenotic, severely stenotic, or totally occluded (see for more details). We performed histological staining with hematoxylin and eosin and Weigert van Gieson and Resorcin-fuchsin (elastica), as well as immunohistochemistry of alpha-smooth muscle (SM) actin and endothelial markers ERG ( D , L , P , X ) or CD31 ( H , T , Z2 ). Magnification was 20×.

Article Snippet: We carried out immunostaining for the expression of alpha-smooth muscle actin (primary antibody: REF: 202M, Cell Marque Corp., a part of MilliporeSigma, Rocklin, CA, USA; secondary antibody: provided as part of the UltraView Universal DAB Detection Kit, REF: 760–500, Ventana Medical Systems, Inc., Roche Group, Tucson, AZ, USA), endothelial marker CD31 (primary antibody: REF: NB100-2284, Novus Biologicals, LLC, Centennial, CO, USA; secondary antibody: REF: BA-1000, Vector Laboratories, Inc., Burlingame, CA, USA), and endothelial marker ERG (primary antibody: REF: 434R, Cell Marque Corp., a part of MilliporeSigma, Rocklin, CA, USA; secondary antibody: provided as part of the OptiView DAB IHC Detection Kit, REF: 760–700, Ventana Medical Systems, Inc., Roche Group, Tucson, AZ, USA).

Techniques: Control, Staining, Immunohistochemistry