cd31 af700  (Novus Biologicals)

Journal: Biofabrication

Article Title: Development of human-derived, three-dimensional respiratory epithelial tissue constructs with perfusable microvasculature on a high-throughput microfluidics screening platform

doi: 10.1088/1758-5090/ac32a5

Figure Lengend Snippet: Immunofluorescence (IF) of alveolar population at the epithelial layer (a) and the epithelial-endothelial junction (c). (a) maximum projection of alveolar layer only. DAPI (blue), phalloidin (red), caveolin-1/CAV1 (cyan), ATP-binding cassette class A3/ABCA3 (yellow). (b) xyz-view of DAPI (blue), CAV1 (cyan), ABCA3 (yellow) from (a). (c) maximum projection of vascularized alveolar tissue construct (contains both epithelial and vascular layers). DAPI (blue), phalloidin (red), CD31 (cyan), ACE2 (yellow). (d) live perfusion of alveoli-capillary construct at timepoint t=0 min. The dye was introduced into the reservoirs of the left parent channel (input channel/vessel) and the white arrow indicatesthe entry point from the vascular network into the output channel/vessel. Taken with Opera Phenix. Scale bar = 100 μm unless otherwise indicated in images.

Article Snippet: To identify the vascular cells comprising the microvascular network in the gel chamber and the side lanes, FITC-conjugated vimentin (eBioscience TM , 11–9897-82) and AF700-conjugated CD31 (R&D Systems, FAB3567N-100) were used.

Techniques: Immunofluorescence, Binding Assay, Construct

Journal: Biofabrication

Article Title: Development of human-derived, three-dimensional respiratory epithelial tissue constructs with perfusable microvasculature on a high-throughput microfluidics screening platform

doi: 10.1088/1758-5090/ac32a5

Figure Lengend Snippet: Immunofluorescence (IF) of small airway epithelial population at the epithelial level (a) and at the epithelial-endothelial junction (c), (d). (a) DAPI (blue), α-tubulin (green), MUC5AC (yellow), CK5 (magenta). (b) xyz-view of DAPI (blue), α-tubulin (green), MUC5AC (yellow), and CK5 (magenta) co-stain to show pseudostratified columnar epithelium structure of the small airway layer. (c) maximum projection of epithelial layer only. α-tubulin (green), ACE2 (magenta) along the surface/higher z-stacks of small airway epithelium. (d) maximum projection at vascular layer. DAPI (blue), CD31 (yellow), ACE2 (magenta) along vasculature layer/lower z-stacks of small airway-capillary constructs. (e) live perfusion of small airway-capillary construct at timepoint t=0 min. The dye was introduced into the reservoirs of the left parent channel (input channel/vessel) and the white arrows indicate the entry point from the vascular network into the output channel/vessel. Taken with Opera Phenix. Scale bar = 100 μm unless otherwise indicated in images.

Article Snippet: To identify the vascular cells comprising the microvascular network in the gel chamber and the side lanes, FITC-conjugated vimentin (eBioscience TM , 11–9897-82) and AF700-conjugated CD31 (R&D Systems, FAB3567N-100) were used.

Techniques: Immunofluorescence, Staining, Construct

Journal: Biofabrication

Article Title: Development of human-derived, three-dimensional respiratory epithelial tissue constructs with perfusable microvasculature on a high-throughput microfluidics screening platform

doi: 10.1088/1758-5090/ac32a5

Figure Lengend Snippet: Flow cytometric evaluation of cell types in 3D respiratory tissue construct and table with cluster frequencies. (a) bivariate plot of AQP5 and Sp-C in 3D alveoli-capillary construct. (b) bivariate plot of AQP5 and Sp-C on transwell platform, alveolar monoculture. (c) bivariate plot of AQP5 and Sp-C in submerged T-75 alveolar monoculture. (d) bivariate plot of a-tubulin and MUC5AC in 3D small airway-capillary construct. (e) bivariate plot of α-tubulin and MUC5AC on transwell platform, small airway monoculture. (f) bivariate plot of α-tubulin and MUC5AC in submerged, T-75 small airway culture. Grey = negative control; pink – stained sample. (g) bivariate plot of CD31 and vimentin in 3D respiratory construct. (h) bivariate plot of CD31 and vimentin of submerged T-75 monoculture for endothelial cells, fibroblasts and pericyte, contour plots of cell types overlaid. (i) Stacked column graph of cell populations from (a)-(f). VAMC = vascularized alveolar multi-chip; VBMC = vascularized bronchiolar multi-chip; AEC = alveolar epithelial cells; SAEC = small airway epithelial cells; TW = transwell. Blue = endothelial cells; red = fibroblasts; black = pericytes. FlowJo for compensation matrix calculation and gating analysis.

Article Snippet: To identify the vascular cells comprising the microvascular network in the gel chamber and the side lanes, FITC-conjugated vimentin (eBioscience TM , 11–9897-82) and AF700-conjugated CD31 (R&D Systems, FAB3567N-100) were used.

Techniques: Construct, Negative Control, Staining

Journal: Frontiers in Immunology

Article Title: Naïve CD4+ T Cell Lymphopenia and Apoptosis in Chronic Hepatitis C Virus Infection Is Driven by the CD31+ Subset and Is Partially Normalized in Direct-Acting Antiviral Treated Persons

doi: 10.3389/fimmu.2021.641230

Figure Lengend Snippet: Naïve CD4+ proportions are lower in HCV infected persons compared to direct-acting antiviral (DAA) -treated persons and age-range matched uninfected controls and the HCV infected persons with cirrhosis have lower naïve CD4+ and CD4+CD31+ proportions compared to those without cirrhosis before and after DAA therapy. In a cross-sectional cohort study; chronic HCV infected persons (n=34), HCV DAA-treated persons at 1-5 years post-DAA therapy initiation (n=29) and age-range matched uninfected controls (n=25) we compared proportions (%) of lymphocyte gated T cells that were naïve CD4 T cells (CD3+CD4+ CD27+CD45RA+) (A) . The HCV infected and DAA-treated groups were stratified by cirrhosis status defined by Transient Elastography scores with cirrhotics: >12.5 kilopaskals (kPa) and non-cirrhotics: <12.5 kPa. Proportions of naïve CD4+ and naïve CD4+CD31+ T cells were assessed (B, C) . Mann Whitney p values shown when p<0.05.

Article Snippet: PBMCs were labeled with anti- CD3-PERCP/CD4-Pacific Blue/CD8-APC-CY7/CD27-AF700/CD45RA-PE-CY7/CD31-BUV-395 (BD Biosciences, San Jose, CA).

Techniques: Infection, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Naïve CD4+ T Cell Lymphopenia and Apoptosis in Chronic Hepatitis C Virus Infection Is Driven by the CD31+ Subset and Is Partially Normalized in Direct-Acting Antiviral Treated Persons

doi: 10.3389/fimmu.2021.641230

Figure Lengend Snippet: BCL-2 expression after IL-7-stimulation was greater in CD31+ and CD31- naïve CD4+ T cell subsets from HCV infected persons compared to naïve CD4+ T cells from HCV DAA-treated persons and uninfected controls, while T cell Receptor-dependent cell cycle entry was similar in naïve CD4+ T cell subsets in all three groups. Magnetic bead purified (negative selection) naïve CD4 T cells from chronic HCV infected (filled circles, n=8), HCV DAA-treated (open circles, n=8) and age-range matched uninfected control (stars, n=7) groups were stimulated with 10ng/ml of recombinant human IL-7 or 1ul anti-CD3/anti-CD28 Activator for 5 days. On 0, 3, and 5 days, flow cytometric analysis of BCL-2 following IL-7 stimulation (A, B) and Ki67 following anti-CD3/anti-CD28 stimulation (C, D) on naïve (CD27+CD45RA+) CD4+CD31+ and CD4+CD31- T cells was performed. Mann Whitney test was used for comparisons between two groups; p= <0.05 considered significant. NS represents non-significant p values.

Article Snippet: PBMCs were labeled with anti- CD3-PERCP/CD4-Pacific Blue/CD8-APC-CY7/CD27-AF700/CD45RA-PE-CY7/CD31-BUV-395 (BD Biosciences, San Jose, CA).

Techniques: Expressing, Infection, Purification, Selection, Recombinant, MANN-WHITNEY