Journal: bioRxiv

Article Title: Endothelial Heterogeneity Across Vascular Beds Impacts Inflammatory Signaling and Neutrophil Adhesion

doi: 10.64898/2026.05.26.727909

Figure Lengend Snippet: Multiplexed ELISA of conditioned media from unstimulated umbilical artery (HUAEC), umbilical vein (HUVEC), dermal microvascular (HDMEC), and pulmonary microvascular (HPMEC) endothelial cells. Conditioned media was collected from 1 well per cell type after 16 hours incubation. Samples were analyzed with a Luminex Magpix device and a custom ProcartaPlex inflammation panel. Data are represented as (A) Heatmap normalized across the range for each protein, with darker tones corresponding to higher expression and less expression for lighter tones. Factors that measured below the limit of detection are indicated with an X. (B) log 10 (average expression) ± SEM for N=4 independent experiments. Factors that measured below the limit of detection are indicated with an X. Full statistical analysis with one-way ANOVA and Tukey-Kramer pairwise comparisons testing was done for average expression across all cell types, with results presented in Supplemental Figure 1.

Article Snippet: The following cells were used: pooled human umbilical vein endothelial cells (HUVEC, PromoCell C-12203, Heidelberg, Germany), human umbilical arterial endothelial cells (HUAEC, PromoCell C-12202, Heidelberg, Germany), adult dermal microvascular endothelial cells (HDMEC, PromoCell C-12212, Heidelberg, Germany), and adult pulmonary microvascular endothelial cells (HPMEC, PromoCell C-12281, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Luminex, Expressing

Journal: bioRxiv

Article Title: Endothelial Heterogeneity Across Vascular Beds Impacts Inflammatory Signaling and Neutrophil Adhesion

doi: 10.64898/2026.05.26.727909

Figure Lengend Snippet: Multiplexed ELISA of conditioned media from TNF-stimulated (50ng/mL TNF in media) umbilical artery (HUAEC), umbilical vein (HUVEC), dermal microvascular (HDMEC), and pulmonary microvascular (HPMEC) endothelial cells. Conditioned media was collected from 1 well per cell type after 16 hours incubation. Samples were analyzed with a Luminex Magpix device and a custom ProcartaPlex inflammation panel. (A) Heatmap normalized across the range for each protein, with darker tones corresponding to higher expression and less expression for lighter tones. (B) Fold-change expression differences from unstimulated control for factors with >8 fold increase by at least one cell type. Data are average fold change-expression for N=4 independent experiments. One-way ANOVA with Tukey-Kramer pairwise comparisons testing was done for each condition, with asterisks indicating statistical significance between conditions (*p<0.05).

Article Snippet: The following cells were used: pooled human umbilical vein endothelial cells (HUVEC, PromoCell C-12203, Heidelberg, Germany), human umbilical arterial endothelial cells (HUAEC, PromoCell C-12202, Heidelberg, Germany), adult dermal microvascular endothelial cells (HDMEC, PromoCell C-12212, Heidelberg, Germany), and adult pulmonary microvascular endothelial cells (HPMEC, PromoCell C-12281, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Luminex, Expressing, Control

Journal: bioRxiv

Article Title: Endothelial Heterogeneity Across Vascular Beds Impacts Inflammatory Signaling and Neutrophil Adhesion

doi: 10.64898/2026.05.26.727909

Figure Lengend Snippet: Multiplexed ELISA of conditioned media from P. aeuriginosa -stimulated (0.05 OD in media) umbilical artery (HUAEC), umbilical vein (HUVEC), dermal microvascular (HDMEC), and pulmonary microvascular (HPMEC) endothelial cells. Conditioned media was collected from 1 well per cell type after 16 hours incubation. Samples were analyzed with a Luminex Magpix device and a custom ProcartaPlex inflammation panel. (A) Heatmap normalized across the range for each protein, with darker tones corresponding to higher expression and less expression for lighter tones. (B) Fold-change expression differences from unstimulated control for factors with >8 fold increase by at least one cell type. Data are average fold change-expression for N=4 independent experiments. One-way ANOVA with Tukey-Kramer pairwise comparisons testing was done for each condition, with asterisks indicating statistical significance between conditions (*p<0.05).

Article Snippet: The following cells were used: pooled human umbilical vein endothelial cells (HUVEC, PromoCell C-12203, Heidelberg, Germany), human umbilical arterial endothelial cells (HUAEC, PromoCell C-12202, Heidelberg, Germany), adult dermal microvascular endothelial cells (HDMEC, PromoCell C-12212, Heidelberg, Germany), and adult pulmonary microvascular endothelial cells (HPMEC, PromoCell C-12281, Heidelberg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Luminex, Expressing, Control

Journal: bioRxiv

Article Title: Endothelial Heterogeneity Across Vascular Beds Impacts Inflammatory Signaling and Neutrophil Adhesion

doi: 10.64898/2026.05.26.727909

Figure Lengend Snippet: Endothelial cells were cultured in 96-well plates and activated with control, TNF(50 ng/mL) or P. aeruginosa (0.05 OD) media for 2 hours. Neutrophils isolated from whole blood were added to each well and incubated for 15 minutes, then imaged before and after 5x PBS washes. (A) Representative images of endothelial cells (red) and neutrophils (green) before and after washing to remove non-adherent cells. (Scale bar = 100 μm) (B, C) Neutrophil adhesion fraction was quantified by counting neutrophils adherent after washing and dividing by the neutrophil count before washing. Data represent average adhesion fraction ± SEM for 3 independent experiments per cell type. One-way ANOVA with Tukey-Kramer pairwise comparisons testing was done for each condition, with asterisks indicating statistical significance between conditions (*p<0.05).

Article Snippet: The following cells were used: pooled human umbilical vein endothelial cells (HUVEC, PromoCell C-12203, Heidelberg, Germany), human umbilical arterial endothelial cells (HUAEC, PromoCell C-12202, Heidelberg, Germany), adult dermal microvascular endothelial cells (HDMEC, PromoCell C-12212, Heidelberg, Germany), and adult pulmonary microvascular endothelial cells (HPMEC, PromoCell C-12281, Heidelberg, Germany).

Techniques: Cell Culture, Control, Isolation, Incubation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Tumor-derived sphingosine-1-phosphate shapes angiogenesis in the acidic microenvironment of osteosarcoma via paracrine and autocrine signaling

doi: 10.3389/fcell.2026.1831997

Figure Lengend Snippet: Acid-stimulated S1P, released by tumor cells, promotes tubulogenesis. (A,B) Quantifications and representative images of HUVEC-GFP cells treated with the CM of OS spheroids cultured in neutral or acidic conditions for 24 h (scale bar: 800 μm). The corresponding control (−) consisted of medium maintained at the same pH as that used for the spheroids. Mann–Whitney test (*p < 0.05 and **p < 0.01 versus neutral control; §§p < 0.01 vs. CM acid control; ###p < 0.005 vs. CM neutral control; n = 12). (C) S1P levels in CM of OS spheroids cultured for 96 h, at nuetral and acidic conditions, with or w/o FTY720 were determined by ELISA. Mann–Whitney test (*p < 0.05 vs. neutral control, #p < 0.05 vs. acid control). (D,E) Quantifications and representative images of HUVEC-GFP cells treated with the CM of OS spheroids, treated with FTY720, as indicated (scale bar: 800 μm). Mann–Whitney test (*p < 0.05 and ***p < 0.005 vs. neutral untreated control; #p < 0.05 vs. CM acid untreated control; n = 12). For all the experiments data are presented as mean ± SEM.

Article Snippet: Human umbilical vein endothelial cells (HUVEC-GFP) were obtained from PromoCell (Heidelberg, Germany) and cultured in Endothelial Cell Basal Medium (ECBM) (PromoCell) supplemented with Endothelial Cell Growth Medium Supplement Pack (PromoCell), 8% Fetal Bovine Serum (FBS) (Lonza, Basil, Switzerland) and 1% Penicillin/Streptomycin (EuroClone, Milan, Italy).

Techniques: Cell Culture, Control, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Tumor-derived sphingosine-1-phosphate shapes angiogenesis in the acidic microenvironment of osteosarcoma via paracrine and autocrine signaling

doi: 10.3389/fcell.2026.1831997

Figure Lengend Snippet: S1P stimulates endothelial cells sprouting. (A) Representative images of endothelial vessel formation after 24 h in a microfluidic platform, shown as whole-channel microscopy (Xpress Pico, upper panels) and high-magnification confocal microscopy (Nikon, lower panels) images. Three-lane Mimetas® microchambers were used, with HUVEC-GFP cells seeded in the top channel, Matrigel® loaded into the central channel, and the bottom channel reserved for the angiogenic cocktail (VEGF with or w/o 250 nM S1P). Dashed boxes highlight regions shown in the confocal close-ups. Confocal images display maximum-intensity projections and 3D side-rendered views. Scale bars: 50 μm. (B) Sprouting quantification (Mann–Whitney test; *p < 0.05 vs. control, n = 10). (C) HUVEC cell proliferation in neutral or acidic medium in monolayer for the indicated time points. Cells were treated with or w/o S1P added in the medium. The total number of cells was assessed by staining of cell nuclei by Hoechst. Data presented as mean ± SEM. Mann–Whitney U test (*p < 0.05 vs. control, n = 8). (D,E) Quantification and representative images of node number and total tubule length in HUVEC-GFP cells, treated with different S1P concentrations (nM), 12 h after seeding. (Mann–Whitney test; *p < 0.05 versus control, n = 9).

Article Snippet: Human umbilical vein endothelial cells (HUVEC-GFP) were obtained from PromoCell (Heidelberg, Germany) and cultured in Endothelial Cell Basal Medium (ECBM) (PromoCell) supplemented with Endothelial Cell Growth Medium Supplement Pack (PromoCell), 8% Fetal Bovine Serum (FBS) (Lonza, Basil, Switzerland) and 1% Penicillin/Streptomycin (EuroClone, Milan, Italy).

Techniques: Microscopy, Confocal Microscopy, MANN-WHITNEY, Control, Staining