bandeiraea simplicifolia conjugated with biotin  (Vector Laboratories)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Fetal growth restriction and placental defects in obese mice are associated with impaired decidualisation: the role of increased leptin signalling modulators SOCS3 and PTPN2

doi: 10.1007/s00018-024-05336-7

Figure Lengend Snippet: Maternal obesity affects fetal-placental growth at embryonic day (E) 18.5. A Number of implantation sites at E6.5 and E18.5 pregnancies in chow-diet (CD) and high-fat diet (HFD) mice. B Representative images of fetuses at E18.5 collected from CD and HFD mice. C Analysis of fetal length and weight at E18.5 and D placental length and weight at E18.5 collected from CD and HFD mice, n indicates number of fetuses or placentas per diet. E Placental efficiency is measured by the fetal-weight-to-placental weight ratio index (FPI). F Haematoxylin and eosin (H&E) of midsagittal sections of E18.5 placentas from CD and HFD mice—labyrinthine zone (LZ) and junctional zone (JZ). G Isolectin BSI-B4 immunohistochemistry of fetal vasculature in the placental labyrinth at E18.5 in placentas collected from CD and HFD mice. H Ratio labyrinth and junctional zone to the placental area at E18.5 in placentas collected from CD and HFD mice, n indicates number of placental sections from 3 to 4 placentas per mother/diet, n = 3 CD and n = 4 HFD. All data are mean ± SEM with individual values from placental sections. Statistical analysis between groups was carried out using Mann–Whitney. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The primary antibodies used were: anti-KI67 (ab15580; Abcam, Cambridge, United Kingdom), anti-PR (D8Q2J; Cell Signaling Technology, Danvers, USA), anti-SOCS3 (ab16030; Abcam), anti-PTPN2 (ab180764; Abcam), anti-pSTAT3 (ab86430; Abcam), Anti-Vimentin (D21H3; Cell Signaling Technology), anti-pan Cytokeratin (C2562, Sigma Aldrich, Saint Louise, USA), anti-CDH1 (610,181, BD Biosciences, New Jersey) and biotin-conjugated isolectin from Bandeiraea simplicifolia BSI-B4 (L2140, Sigma).

Techniques: Immunohistochemistry, MANN-WHITNEY

Journal: Journal of Tissue Engineering

Article Title: Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold

doi: 10.1177/20417314221140979

Figure Lengend Snippet: hLEC seeded scaffolds in vivo. (a) Low magnification image of section through an entire hLEC seeded scaffold (demarcated by a blue line, with the skin above) at 7 days in vivo. Area within the blue rectangle appears at higher magnification in (b) and (c) and includes numerous hLEC vessels positive for hCD31/podoplanin. Black arrows at the edge of the scaffold indicate rat tissue infiltration into the pores of the scaffold. * indicates scaffold material (appears white). Scale bar = 1000 µm. (b) Area within the blue rectangle in (a) showing numerous hLEC capillaries (arrows) immunostained positive for hCD31. The hLEC capillaries are surrounded by rat infiltrating connective tissue. * scaffold material. Scale bar = 200 µm. (c) Area within the blue rectangle of (a) and adjacent section to (b) with the same hLEC capillaries strongly immunostained with podoplanin (arrows). * indicates scaffold material. Scale bar = 200 µm. (d) Intermingling of hLEC capillaries (stained brown with hCD31, black arrow) and rat tissue in vivo at 7 days. Rat vessels stain purple (blue arrow) with Griffonia simplicifolia lectin. Scale bar = 100 µm. (e) Mosaic vessel in an hLEC seeded scaffold at 7 days in vivo. The black arrow (lower right) indicates a hCD31 + hLEC vessel (brown) joined to a mosaic vessel that has hCD31 + ECs (brown, indicated by black arrow at top) and rat mesenchymal support cells (purple, indicated by blue arrow) wrapping around its abluminal surface. Rat blood is seen in the vessel lumen. Scale bar = 100 µm. (f) Numerous podoplanin + hLEC capillaries (arrows) in a scaffold in vivo at 14 days. Some human lymphatics show white blood cell only trafficking in their lumens. Scale bar = 100 µm. (g) hLEC + hvSMC seeded scaffold at 14 days in vivo demonstrating lymphatic function. Podoplanin + human lymphatic vessel (arrow) exclusively trafficking rat white blood cells within its lumen. Scale bar = 50 µm. (h) Low magnification image of hLEC + hvSMC seeded scaffold at 14 days in vivo. Arrow indicates HG19 + rat lymphatic positioned centrally in the scaffold. Scale bar = 200 µm. (i) The area in the blue box in (h) demonstrating the rat lymphatic vessel (HG19 + , black arrow) positioned centrally in a hLEC + hvSMC seeded scaffold at 14 days in vivo. The thick dotted black arrow indicates a branch point from the rat lymphatic. The blue arrow indicates the branch which is not HG19 + . Scale bar = 50 µm. (j) Adjacent tissue section to (i) where the original HG19 + rat lymphatic is seen (black arrow) and the blue arrow indicates a human podoplanin + lymphatic vessel, indicating inosculation between rat and human lymphatics within the scaffold. Scale bar = 50 µm.

Article Snippet: Sections were washed and overlaid with Griffonia simplicifolia lectin-biotin conjugate (1:300, Vector Laboratories) for 30 min, followed by detection with Streptavidin-HRP conjugate (30 min, 1:400, Dako) and Vector VIP substrate kit (5 min, Vector Laboratories).

Techniques: In Vivo, Staining

Journal: Journal of Tissue Engineering

Article Title: Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold

doi: 10.1177/20417314221140979

Figure Lengend Snippet: hBEC seeded scaffolds in vivo. (a) Low magnification image of hBEC seeded scaffold (demarcated by the blue line) in the skin at 7 days in vivo. Black arrow indicates hBEC immuno-stained positive for hCD31. Blue arrows indicate rat connective tissue infiltration into the scaffold pores from the rat skin above the scaffold. * indicates scaffold material (white). Scale bar = 1000 µm. (b) Higher magnification image of hCD31 + hBEC capillaries (arrows) at 7 days in vivo. The grey material around the hBEC capillaries is human fibrin. Scale bar = 200 µm. (c) Low magnification image of hBEC + hvSMC seeded scaffold (demarcated by the blue line) in a skin wound at 7 days in vivo. Black arrows indicate human capillaries immuno-stained positive with hCD31. These human vessels appear more numerous than those in (a). Blue arrows indicate rat connective tissue infiltration into the scaffold pores from the rat skin above the scaffold. * indicates scaffold material (white). Scale bar = 1000 µm. (d) Higher magnification image of hCD31 + hBEC + hvSMC capillaries at 7 days in vivo. The grey material around the capillaries is human fibrin. The human capillaries (arrows) appear larger than in (b) and contain rat blood. * indicates scaffold material. Scale bar = 200 µm. (e) Intermingling of human hCD31 + hBEC capillaries (immuno-stained brown, black dashed arrow) with rat capillaries (stained with Griffonia simplicifolia lectin (purple), blue dashed arrow). The vessels appear to contact each other at the thick black arrow. Scale bar = 50 µm. (f, g) Adjacent tissue sections through a hBEC + hvSMC seeded scaffold at 7 days in vivo. (f) is immuno-stained for hCD31, numerous positive human capillaries are seen (brown, arrows). In (g) the same capillaries as (f) are immuno-stained for podoplanin, but are negative (arrows). Scale bars = 100 µm. (h) FITC dextran tail vein infusion of nude rat at 7 days. hCD31 + hBEC capillaries (red, white dashed arrows) contain FITC dextran (green, white solid arrows) indicating inosculation between human and rat capillaries in the scaffold. Scale bar = 50 µm.

Article Snippet: Sections were washed and overlaid with Griffonia simplicifolia lectin-biotin conjugate (1:300, Vector Laboratories) for 30 min, followed by detection with Streptavidin-HRP conjugate (30 min, 1:400, Dako) and Vector VIP substrate kit (5 min, Vector Laboratories).

Techniques: In Vivo, Staining