human umbilical vein endothelial cells huvec gfp  (PromoCell)

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

Journal: bioRxiv

Article Title: Metabolic Reprogramming of Human Macrophages Drives the Formation of Hybrid M1/M2 Pro-Regenerative Extracellular Vesicles

doi: 10.64898/2026.01.16.699890

Figure Lengend Snippet: (A) Quantitative analysis of BrdU incorporation assay measuring endothelial cell proliferation. (B) Quantitative analysis of HUVEC migration and (C) representative images of HUVEC migration. Scale bar=100 µm. (D) Plot of HUVEC migration for 24 hours. (E) Representative images of GFP-HUVECs which were treated with 1 µg/mL EVs for 9 hours on GelTrex. Scale bar=500 µm. (F) Quantitative analysis of number of branches and (G) total tube length. Error bars in the graphs represent mean ± SD of EVs obtained from three or four independent donors. ****p < 0.0001, ***p <0.001, **p < 0.01, *p <0.05 (one-way ANOVA and Tukey multiple comparison).

Article Snippet: Human umbilical vein endothelial cells (HUVECs) (Promocell, C-12203) and Green Fluorescent Protein (GFP) -HUVECs (Angio-proteomie, cAP-0001GFP) were cultured in endothelial cell basal medium (EBM) plus endothelial cell growth medium supplements (Promocell, C-22011) supplemented with 10% FBS (Gibco) and 1% Penicillin/Streptomycin (100 U/mL, Sigma, P4333) and incubated at 37°C in a humified 5% CO 2 atmosphere.

Techniques: BrdU Incorporation Assay, Migration, Comparison

Journal: bioRxiv

Article Title: Metabolic Reprogramming of Human Macrophages Drives the Formation of Hybrid M1/M2 Pro-Regenerative Extracellular Vesicles

doi: 10.64898/2026.01.16.699890

Figure Lengend Snippet: (A) Quantitative analysis of BrdU incorporation assay measuring endothelial cell proliferation. (B) Quantitative analysis of HUVEC migration and (C) representative images of HUVEC migration. Scale bar= 100 µm. (D) Representative images of GFP-HUVECs which were treated with 1 µg/mL EVs for 9 hours on GelTrex. Scale bar=500 µm. ( E ) Quantitative analysis of number of branches and ( F ) total tube length. (G) Quantitative analysis of BrdU incorporation assay measuring MSC proliferation. (H) Quantitative analysis of MSC migration. (I) Alkaline phosphatase activity at day 7. (J) Alizarin Red staining at day 14 for mineralized matrix deposition (Scale bar=100 µm) and (K) quantification of calcium content. Error bars in the graphs represent mean ± SD of EVs obtained from three or four independent donors. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 (one-way ANOVA and Tukey multiple comparison). Error bars in the graphs represent mean ± SD of EVs obtained from three or four independent donors.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) (Promocell, C-12203) and Green Fluorescent Protein (GFP) -HUVECs (Angio-proteomie, cAP-0001GFP) were cultured in endothelial cell basal medium (EBM) plus endothelial cell growth medium supplements (Promocell, C-22011) supplemented with 10% FBS (Gibco) and 1% Penicillin/Streptomycin (100 U/mL, Sigma, P4333) and incubated at 37°C in a humified 5% CO 2 atmosphere.

Techniques: BrdU Incorporation Assay, Migration, Activity Assay, Staining, Comparison

Journal: Cancer immunology research

Article Title: Secretion of a VEGF-blocking scFv enhances CAR T-cell potency

doi: 10.1158/2326-6066.CIR-24-0876

Figure Lengend Snippet: A and B , Bioluminescence of HEK-293T VEGFR reporter cells treated with supernatant containing either anti-VEGF or anti-CD19 and recombinant VEGF (rVEGF) (Data from technical triplicates, points represent the mean, error bars represent SEM, p-value by two-way ANOVA) ( A ) or supernatant from confluent VEGF-producing 786O tumor cells (data from technical triplicates, lines represent the median, p-value by one-way ANOVA) ( B ). C, Schematic of in vitro angiogenesis assay to visualize blood vessel formation upon blocking VEGF. GFP + human umbilical vein endothelial cells (HUVECs) were cocultured with normal human dermal fibroblasts (NHDFs), suramin, rVEGF, and culture supernatant from CAR αCD19 or CAR αVEGF constructs and primitive blood vessel formation was imaged over time on the incucyte. D, Representative images of GFP + HUVECs with vessels labeled by the automated incucyte angiogenesis software package. E, HUVEC blood vessel network length (length of blood vessel network (mm) per mm 2 in image). (data representative of 2 technical replicates, mean±SEM, p-value by two-way ANOVA).

Article Snippet: After 3 days, Human Umbilical Vein Endothelial Cells (HUVECs) (Angio Proteomie, CAP0001GFP) were seeded onto the confluent layer of NHDFs at 5,000 cells/well and treatment conditions were added (recombinant VEGF at 4 ng/ml (R&D Systems 293-VE-010), suramin sodium salt at 0.25 mg/ml (Millipore Sigma, S2671–25MG), culture supernatant from confluent 70 αVEGF , 70 aCD19 , or untransduced Jurkat T cells diluted 1:4).

Techniques: In Vitro, Recombinant, Angiogenesis Assay, Blocking Assay, Construct, Labeling, Software