Journal: Journal of Tissue Engineering

Article Title: Development and characterisation of a low-concentration sodium dodecyl sulphate decellularised porcine dermis

doi: 10.1177/2041731417724011

Figure Lengend Snippet: Images of histological sections of cellular porcine skin, decellularised porcine dermis and decellularised human dermis stained with anti-collagen IV, anti-laminin and GSL-1 isolectin B 4 . The images show positive collagen IV staining (c4) in cellular porcine skin and decellularised human dermis but not in decellularised porcine dermis. Positive laminin staining (lam) was visible in cellular porcine skin, decellularised porcine dermis and decellularised human dermis. Residual alpha-gal remained present throughout the dermis in decellularsied porcine dermis. Small areas of focal staining (lec) were visible in decellularised human dermis. The controls (bottom panel) are labelled with isotype control antibodies (collagen IV and laminin) and galactose blocked GSL-1 isolectin B 4 . Images of anti-collagen IV stained sections captured at 30× magnification, with scale bar 50 μm. All other images captured at 20× magnification, with scale bar 100 μm.

Article Snippet: The alpha-gal epitope was labelled using biotinylated GSL-1 – isolectin B 4 (GSL-1 Lectin; Vector laboratories).

Techniques: Staining

Journal: Journal of tissue engineering and regenerative medicine

Article Title: The fabrication and characterization of a multi-laminate, angle-ply collagen patch for annulus fibrosus repair

doi: 10.1002/term.2250

Figure Lengend Snippet: Representative histological images of (A) fresh and (B) decellularized porcine pericardium stained with haematoxylin and eosin (H&E), respectively (pink = extracellular matrix; arrowheads = location of cell nuclei; total magnification: 200 ×). Representative histological images of (C) fresh porcine pericardium demonstrating positive (brown) immunohistochemistry (IHC) staining for alphagal epitope and (D) decellularized pericardium illustrating a lack of positive staining (blue = matrix; black = cell nuclei; total magnification: 200 ×; inserts = negative IHC controls). (E) Ethidium bromide-stained agarose gels for DNA isolated from fresh (lanes 1–5) and decellularized (lanes 6–10) pericardium (white bands = presence of DNA). A 300–24 000-bp DNA standard ladder (lanes 11–12) is shown for comparison. (F) DNA quantification of fresh and decellularized pericardium performed with Nanodrop spectrophotometry. (G) Diagrammatic representation of multi-laminate annulus fibrosus (AF) patch formation in which at least three-plys of decellularized pericardium were stacked (red tubes = aligned collagen type I fibres in the fibrous pericardium layer of each ply) oriented at ±30° to each other. (H) Schematic representation depicting histology sectioning of AF patches using an oblique cut (dotted red line) across multiple layers performed to microscopically visualize collagen fibre alignment in fibrous pericardium surfaces stacked directly adjacent to one another demonstrating a ±30° ‘chevron’ (*) pattern. Alternatively, when fibrous and parietal pericardium surfaces were directly adjacent to each other, a ‘half chevron’ (#) was achieved. (I) Chevron and (J) half chevron patterns (dashed white outlines), respectively, were observed within each patch via polarized light microscopy confirming the presence of oriented collagen fibre alignment (total magnification: 100 ×). (K) Macroscopic image of a six-ply AF patch sewn with suture (black outline). Solid lines connecting different study groups on the graph indicate a significant difference (p < 0.05)

Article Snippet: Additionally, immunohistochemistry (IHC) for the porcine antigenic epitope alpha-gal (biotinylated GSL1-isolectin B4; 2.5 μg/ml; VectorLabs) was performed.

Techniques: Staining, Immunohistochemistry, Isolation, Spectrophotometry, Light Microscopy

Journal: British Journal of Cancer

Article Title: Inhibition of epidermoid carcinoma A431 cell growth and angiogenesis in nude mice by early and late treatment with a novel dextran derivative

doi: 10.1038/sj.bjc.6600985

Figure Lengend Snippet: Effects of NaPaC on A431 tumour microvessel network. Endothelial cells were stained in early ( A ) and late ( C ) treatment controls, and in early ( B ) and late ( D ) NaPaC-treated tumours using GSL-1 lectin. Microvessel lumens in panels were indicated with asterisks. Magnification used was × 250. The representative AEC-stained endothelial cells (red) are indicated with arrows.

Article Snippet: Endothelial cells were specifically labelled with GSL-1 isolectin B4 (Vector Laboratories, Burlingame, CA, USA).

Techniques: Staining

Journal: British Journal of Cancer

Article Title: Inhibition of epidermoid carcinoma A431 cell growth and angiogenesis in nude mice by early and late treatment with a novel dextran derivative

doi: 10.1038/sj.bjc.6600985

Figure Lengend Snippet: Quantification of endothelial cell density and vessel area in early and late NaPaC-treated tumours. ( A ) The GSL-1 lectin-stained endothelial cells per mm 2 of tumour area (endothelial cell density) and ( B ) the fraction of the total tissue area occupied by the wall or/and lumen (vessel area) was determined as described in Materials and Methods. Each column represents the mean ± s.d. ( n =10). * P <0.05 vs control.

Article Snippet: Endothelial cells were specifically labelled with GSL-1 isolectin B4 (Vector Laboratories, Burlingame, CA, USA).

Techniques: Staining

Journal: Journal of Tissue Engineering

Article Title: Development and characterisation of a low-concentration sodium dodecyl sulphate decellularised porcine dermis

doi: 10.1177/2041731417724011

Figure Lengend Snippet: Images of histological sections of cellular porcine skin, decellularised porcine dermis and decellularised human dermis stained with anti-collagen IV, anti-laminin and GSL-1 isolectin B 4 . The images show positive collagen IV staining (c4) in cellular porcine skin and decellularised human dermis but not in decellularised porcine dermis. Positive laminin staining (lam) was visible in cellular porcine skin, decellularised porcine dermis and decellularised human dermis. Residual alpha-gal remained present throughout the dermis in decellularsied porcine dermis. Small areas of focal staining (lec) were visible in decellularised human dermis. The controls (bottom panel) are labelled with isotype control antibodies (collagen IV and laminin) and galactose blocked GSL-1 isolectin B 4 . Images of anti-collagen IV stained sections captured at 30× magnification, with scale bar 50 μm. All other images captured at 20× magnification, with scale bar 100 μm.

Article Snippet: The alpha-gal epitope was labelled using biotinylated GSL-1 – isolectin B 4 (GSL-1 Lectin; Vector laboratories).

Techniques: Staining

Journal: Nature Communications

Article Title: Fluorinated rhamnosides inhibit cellular fucosylation

doi: 10.1038/s41467-021-27355-9

Figure Lengend Snippet: a Effect of 10 or 100 µM Fucotrim I ( 9 ) & II ( 10 ) and P-Fuc2F on cell surface glycosylation using different lectins, presented as percentage lectin binding compared to DMSO control in a Box and Whisker Plot with 5–95 percentiles (Supplementary Fig. ). N ≥ 3 biologically independent experiments of 10.000 gated cells per sample and only N = 2 technical replicates per experiment for DMSO control. ( b ) Effect of C6-modifications on potency of inhibition presented as mean ± SEM. N ≥ 3 biologically independent experiments of 10.000 gated cells per sample and N = 2 technical replicates for each experiment. c Potency of Fucotrim I ( 9 ) and II ( 10 ) (blue) compared to known salvage pathway-dependent fucose inhibitors P-Fuc2F, Fucostatin-I and Fucostatin-II (red) using AOL lectin ( n = 3) presented as mean ± SEM. N ≥ 3 biologically independent experiments of 10.000 gated cells per sample and N = 2 technical replicates for each experiment. d Onset and recovery of defucosylation. THP1 cells were incubated with 10 or 100 µM (Supplementary Fig. ) compound or DMSO control and fucosylation levels were determined with AAL lectins or AOL lectins (Supplementary Fig. ) for 6 days, presented as mean ± SEM. N ≥ 3 biologically independent experiments of 10.000 gated cells per sample and N = 2 technical replicates for each experiment. e The effect on viability (blue) and cytotoxicity (red) on THP1 cells after incubation with 2, 10, 100, or 250 µM 6 – 10 or 1 – 5 (Supplementary Fig. ) for 3 days was determined with an XTT and LDH respectively and presented as percentage cell viability or cytotoxicity compared to DMSO control, presented as mean ± SEM. N = 1 biologically independent experiments of 10.000 gated cells per sample and N = 2 technical replicates.

Article Snippet: Biotinylated AAL, SNA, MAL-II, WGA, LCA, PSA, PNA, PHA-L, and GSL-1 lectins and 10x Carbo-free Blocking Buffer were purchased from Vector laboratories Inc. Biotinylated AOL lectin was purchased from TCI Europe.

Techniques: Binding Assay, Whisker Assay, Inhibition, Incubation