sheep anti human cd31 pecam 1  (R&D Systems)

Journal: bioRxiv

Article Title: Paracrine Factor Local Gradient-Generating System for Engineering Perfusable Vascularized Hepatocyte Tissues with Perfusion-Induced Proliferation

doi: 10.1101/2025.11.10.687539

Figure Lengend Snippet: A: Schematic illustrations of PFLG generated by hLF cryogel, inducing microvessel penetration into hepatocyte tissue. B: Fluorescent projection images of vascularized hepatocyte tissue on day 5. Blue: cell nuclei (Hoechst 33342), green: F-actin (Alexa Fluor 488-phalloidin), red: HUVECs (CD31). Arrowheads indicate microvessels penetrating hepatocyte tissue. Asterisks in the magnified image indicate dead cells in hepatocyte tissue. Scale bars, 200 µm (upper panel); 50 µm (lower panel, magnified images). C: Quantitative analysis of vessel area ratio in fluorescent projection images of vascularized hepatocyte tissues on day 5. D: Quantitative analysis of the number of penetrating vessels on day 5. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (One-way ANOVA followed by Tukey’s HSD test).

Article Snippet: Samples were then incubated overnight at 4°C with the following primary antibodies: sheep anti-PECAM-1 (CD31) (AF806, R&D Systems, Minneapolis, MN, USA) to label HUVECs; mouse anti-multidrug resistance-associated protein 2 (MRP2) (NBP1-42349, Novus Biologicals, Centennial, CO, USA) to visualize bile canaliculi formed by rHeps; and rabbit anti-Ki67 (ab16667, Abcam, Cambridge, UK) to identify proliferating cells.

Techniques: Generated

Journal: bioRxiv

Article Title: Paracrine Factor Local Gradient-Generating System for Engineering Perfusable Vascularized Hepatocyte Tissues with Perfusion-Induced Proliferation

doi: 10.1101/2025.11.10.687539

Figure Lengend Snippet: A: Immunofluorescence confocal images of vascularized hepatocyte tissue (rHep:HUVEC = 9:1) on day 5. Blue: cell nuclei (Hoechst 33342), green: F-actin (Alexa Fluor 488-phalloidin), red: HUVECs (CD31). B: Cross-sectional images of vascularized hepatocyte tissue. Arrowheads indicate microvessels penetrating the hepatocyte tissue. The asterisk in the magnified XZ section highlights hepatic sinusoid-like structures, where hepatocytes are in close contact with microvessels. Scale bar, 50 µm. C: Schematic illustrations of hepatic sinusoids (H: hepatocytes, D: space of Disse, SEC: sinusoid endothelial cells, S: hepatic sinusoid).

Article Snippet: Samples were then incubated overnight at 4°C with the following primary antibodies: sheep anti-PECAM-1 (CD31) (AF806, R&D Systems, Minneapolis, MN, USA) to label HUVECs; mouse anti-multidrug resistance-associated protein 2 (MRP2) (NBP1-42349, Novus Biologicals, Centennial, CO, USA) to visualize bile canaliculi formed by rHeps; and rabbit anti-Ki67 (ab16667, Abcam, Cambridge, UK) to identify proliferating cells.

Techniques: Immunofluorescence

Journal: bioRxiv

Article Title: Paracrine Factor Local Gradient-Generating System for Engineering Perfusable Vascularized Hepatocyte Tissues with Perfusion-Induced Proliferation

doi: 10.1101/2025.11.10.687539

Figure Lengend Snippet: A: Immunofluorescence projection images of bile canalicular formation in vascularized hepatocyte tissue. Projection images were constructed from Z-stack confocal images. Blue: cell nuclei (Hoechst 33342), green: F-actin (Alexa Fluor 488-phalloidin), red: HUVECs (CD31), pink: bile canaliculi (MRP2). Arrowheads indicate MRP2-positive bile canaliculi. B: Excretion of CellTracker Red CMTPX by polarized hepatocytes in vascularized hepatocyte tissue (red: CMTPX; green: GFP-rHep). Arrowheads indicate CMTPX excreted into bile canaliculi. C: Cross-sectional image showing CMTPX excretion into bile canaliculi in vascularized hepatocyte tissue (red: CMTPX; green: GFP-rHep). Arrowheads indicate CMTPX in bile canaliculi. Scale bars, 50 µm. D: Quantitative analysis of MRP2-positive area ratio in hepatocyte tissue. E: Quantification of albumin secretion from hepatocyte tissues. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (One-way ANOVA followed by Tukey’s HSD test).

Article Snippet: Samples were then incubated overnight at 4°C with the following primary antibodies: sheep anti-PECAM-1 (CD31) (AF806, R&D Systems, Minneapolis, MN, USA) to label HUVECs; mouse anti-multidrug resistance-associated protein 2 (MRP2) (NBP1-42349, Novus Biologicals, Centennial, CO, USA) to visualize bile canaliculi formed by rHeps; and rabbit anti-Ki67 (ab16667, Abcam, Cambridge, UK) to identify proliferating cells.

Techniques: Immunofluorescence, Construct

Journal: bioRxiv

Article Title: Paracrine Factor Local Gradient-Generating System for Engineering Perfusable Vascularized Hepatocyte Tissues with Perfusion-Induced Proliferation

doi: 10.1101/2025.11.10.687539

Figure Lengend Snippet: A: Time-lapse images of fluorescently labeled microbeads (diameter: 1 µm) perfused through vascularized hepatocyte tissue. Arrowheads indicate a perfused microbead, and arrows indicate the flow direction within the microvessel. Scale bar, 50 µm. B: Time-lapse and projection images showing perfusion of 70 kDa FITC-dextran solution through vascularized hepatocyte tissue. Green: FITC-dextran and F-actin (Alexa Fluor 488-phallidin), red: microvessels (CD31). Scale bars, 200 µm (upper panel); 100 µm (lower panel, magnified images). C: Quantitative analysis of cumulative FITC-dextran fluorescence in regions H1–H5 and V1–V5 shown in the right image. Boxes in the projection image indicate regions of interest for analyzing cumulative FITC-dextran fluorescence during the perfusion process (H1–H5: hepatocyte tissue regions; V1–V5: hydrogel regions)

Article Snippet: Samples were then incubated overnight at 4°C with the following primary antibodies: sheep anti-PECAM-1 (CD31) (AF806, R&D Systems, Minneapolis, MN, USA) to label HUVECs; mouse anti-multidrug resistance-associated protein 2 (MRP2) (NBP1-42349, Novus Biologicals, Centennial, CO, USA) to visualize bile canaliculi formed by rHeps; and rabbit anti-Ki67 (ab16667, Abcam, Cambridge, UK) to identify proliferating cells.

Techniques: Labeling, Fluorescence

Journal: bioRxiv

Article Title: Paracrine Factor Local Gradient-Generating System for Engineering Perfusable Vascularized Hepatocyte Tissues with Perfusion-Induced Proliferation

doi: 10.1101/2025.11.10.687539

Figure Lengend Snippet: A: Schematic illustrations of static and perfusion culture conditions. Cells were cultured with hLF cryogels (hLF(+)) until day 3. From day 3 to day 7, some cells were cultured without hLF cryogels (hLF(−)) under either static (0 mmH 2 O) or perfusion (1 or 2 mmH 2 O) conditions. B: Immunofluorescence projection images of vascularized hepatocyte tissue under static and perfusion culture. Projection images were constructed from Z-stack confocal images. Blue: cell nuclei (Hoechst 33342), green: F-actin (Alexa Fluor 488-phalloidin), red: HUVECs (CD31), yellow: proliferating cell nuclei (Ki67). Arrowheads indicate Ki67-positive proliferating hepatocytes. Asterisks indicate bile canaliculi-like structures, showing double-layered F-actin between hepatocytes. Scale bars, 50 µm. C: Representative immunofluorescence images showing Ki67-positive rHeps undergoing mitosis. Arrowheads indicate both interphase nuclei and mitotic chromosomal dynamics. Blue: cell nuclei (Hoechst 33342), green: F-actin (Alexa Fluor 488-phalloidin), red: HUVECs (CD31), yellow: proliferating cell nuclei (Ki67). D: Quantification of the ratio of Ki67-positive rHeps. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (One-way ANOVA followed by Tukey’s HSD test).

Article Snippet: Samples were then incubated overnight at 4°C with the following primary antibodies: sheep anti-PECAM-1 (CD31) (AF806, R&D Systems, Minneapolis, MN, USA) to label HUVECs; mouse anti-multidrug resistance-associated protein 2 (MRP2) (NBP1-42349, Novus Biologicals, Centennial, CO, USA) to visualize bile canaliculi formed by rHeps; and rabbit anti-Ki67 (ab16667, Abcam, Cambridge, UK) to identify proliferating cells.

Techniques: Cell Culture, Immunofluorescence, Construct

Journal: Animal Models and Experimental Medicine

Article Title: Bilateral carotid‐jugular arteriovenous graft implantation in an ovine model is safe and durable for facilitation of arteriovenous graft innovation

doi: 10.1002/ame2.70096

Figure Lengend Snippet: Representative imaging of explanted AV grafts. The images include scanning electron microscopy (SEM) in the first column, immunohistochemisty (IHC) in the second column, and hematoxylin and eosin (H&E) staining in the third column. The IHC imaging includes DAPI staining (dark blue) for cell nuclei and CD31 staining (magenta) for endothelial cell junctions, which when observed in the characteristic circular ring‐like staining pattern as seen here, suggests the presence of endothelial cells. Column 3 demonstrates patent sections of graft on the left and occluded sections of the graft on the right. The image on the far‐right side of the figure demonstrates the graft sectioning technique relative to its orientation with “1” corresponding to the venous anastomosis, “2” corresponding to graft mid‐segment, and “3” corresponding to the arterial anastomosis.

Article Snippet: Samples were then incubated with mouse anti‐sheep CD31 antibody (NB100‐65900, Novus Biologicals; 1:50 in 1% BSA/PBS) at 4°C overnight.

Techniques: Imaging, Electron Microscopy, Staining