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PromoCell human umbilical vein endothelial cells huvecs
Human Umbilical Vein Endothelial Cells Huvecs, supplied by PromoCell, used in various techniques. Bioz Stars score: 98/100, based on 1562 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/huvec-c+human+umbilical+vein+endothelial+cells/pm42243163-46-0-13?v=PromoCell
Average 98 stars, based on 1562 article reviews

human umbilical vein endothelial cells huvecs - by Bioz Stars, 2026-07

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Biocompatibility of PC capsules: (A–B) cell viability of HUVECs (A) and NIH/3T3 cells (B) treated with different concentrations of PC capsules for 24 h detected by MTT assay; (C–D) toxic staining of PC capsules on HUVECs and NIH/3T3 cells detected by living/dead cell staining; (E) photos of the backs of mice after receiving a subcutaneous injection of PC capsules and subcutaneous PC capsules after injection at day 4 and day 7; (F) slices of major organs (heart, liver, spleen, lungs, and kidneys) in mice after 14 days of PC capsule administration; scale bar = 100 μm. Serum liver function indicators were: (G) ALT and (H) AST. Serum renal function indicators were: (I) BUN and (J) CRE. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ns (p > 0.05).

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: Biocompatibility of PC capsules: (A–B) cell viability of HUVECs (A) and NIH/3T3 cells (B) treated with different concentrations of PC capsules for 24 h detected by MTT assay; (C–D) toxic staining of PC capsules on HUVECs and NIH/3T3 cells detected by living/dead cell staining; (E) photos of the backs of mice after receiving a subcutaneous injection of PC capsules and subcutaneous PC capsules after injection at day 4 and day 7; (F) slices of major organs (heart, liver, spleen, lungs, and kidneys) in mice after 14 days of PC capsule administration; scale bar = 100 μm. Serum liver function indicators were: (G) ALT and (H) AST. Serum renal function indicators were: (I) BUN and (J) CRE. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ns (p > 0.05).

Article Snippet: Human umbilical vein endothelial cells (HUVECs) (CRL-1730, USA, RRID: CVCL_2959) and NIH/3T3 cells (CRL-1658, USA, RRID: CVCL_0594) used in this study were purchased from American Type Culture Collection.

Techniques: Capsules, MTT Assay, Staining, Injection, Standard Deviation

PC capsules increased cell viability under H 2 O 2 stimulation: cell viability of HUVECs (A) and NIH/3T3 (B) cells treated with different concentrations of H 2 O 2 for 6 h detected by MTT assay; viability of H 2 O 2 ‐treated HUVECs (C) and NIH/3T3 cells (D) with different concentrations of PC capsule pretreatment. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ∗∗∗∗p < 0.0001.

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: PC capsules increased cell viability under H 2 O 2 stimulation: cell viability of HUVECs (A) and NIH/3T3 (B) cells treated with different concentrations of H 2 O 2 for 6 h detected by MTT assay; viability of H 2 O 2 ‐treated HUVECs (C) and NIH/3T3 cells (D) with different concentrations of PC capsule pretreatment. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ∗∗∗∗p < 0.0001.

Techniques: Capsules, MTT Assay, Standard Deviation

PC capsules restored cell function in H 2 O 2 -treated HUVECs and NIH/3T3 cells. HUVECs and NIH/3T3 cells were treated with PC capsules for 24 h and H 2 O 2 for 6 h. (A) Images of HUVEC migration exposure to H 2 O 2 , NAC, PC capsules after 24 h; (B) quantification of percentage wound area remaining for HUVECs; (C) images of NIH/3T3 cell migration exposure to H 2 O 2 , NAC, PC capsules after 24 h; (D) quantification of percentage wound area remaining for NIH/3T3 cells; (E–F) digital images of endothelial cell microtubule formation after treatment with H 2 O 2 , NAC, and PC capsules. Quantification of (G) number of branch sites, (H) total lengths and (I) numbers of tube nodes. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs. H 2 O 2 group).

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: PC capsules restored cell function in H 2 O 2 -treated HUVECs and NIH/3T3 cells. HUVECs and NIH/3T3 cells were treated with PC capsules for 24 h and H 2 O 2 for 6 h. (A) Images of HUVEC migration exposure to H 2 O 2 , NAC, PC capsules after 24 h; (B) quantification of percentage wound area remaining for HUVECs; (C) images of NIH/3T3 cell migration exposure to H 2 O 2 , NAC, PC capsules after 24 h; (D) quantification of percentage wound area remaining for NIH/3T3 cells; (E–F) digital images of endothelial cell microtubule formation after treatment with H 2 O 2 , NAC, and PC capsules. Quantification of (G) number of branch sites, (H) total lengths and (I) numbers of tube nodes. Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs. H 2 O 2 group).

Techniques: Capsules, Cell Function Assay, Migration, Standard Deviation, Control

PC capsules improved mitochondrial function in H 2 O 2 -treated HUVECs and NIH/3T3 cells: (A–D) elimination of ROS from HUVECs (A, C) and NIH/3T3 (B, D) cells by PC capsules as determined by MitoSOX; (E–H) assessment of PC-mediated HUVECs (E, G) and NIH/3T3 cells (F, H) MMP using TMRM assays; (I–J) effects of PC capsules on ATP production in HUVECs (I) and NIH/3T3 cells (J). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001, ####p < 0.0001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (vs. H 2 O 2 group).

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: PC capsules improved mitochondrial function in H 2 O 2 -treated HUVECs and NIH/3T3 cells: (A–D) elimination of ROS from HUVECs (A, C) and NIH/3T3 (B, D) cells by PC capsules as determined by MitoSOX; (E–H) assessment of PC-mediated HUVECs (E, G) and NIH/3T3 cells (F, H) MMP using TMRM assays; (I–J) effects of PC capsules on ATP production in HUVECs (I) and NIH/3T3 cells (J). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001, ####p < 0.0001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (vs. H 2 O 2 group).

Techniques: Capsules, Standard Deviation, Control

PC capsule–mediated cytoprotection via promoting activation of PI3K/AKT signaling pathway: expression of p-PI3K, PI3K, p-AKT, and AKT in HUVECs treated with PC capsules for 24 h and presence/absence of H 2 O 2 for 6 h (A); and analysis of optical density values of p-PI3K/PI3K (B) and p-AKT/AKT (C); expression of p-PI3K, PI3K, p-AKT, and AKT in NIH/3T3 cells treated with PC capsules for 24 h and presence/absence of H 2 O 2 for 6 h (D); and analysis of optical density values of p-PI3K/PI3K (E) and p-AKT/AKT (F). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: #p < 0.05, ##p < 0.01, ###p < 0.001 (vs control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs PC group or vs H 2 O 2 group).

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: PC capsule–mediated cytoprotection via promoting activation of PI3K/AKT signaling pathway: expression of p-PI3K, PI3K, p-AKT, and AKT in HUVECs treated with PC capsules for 24 h and presence/absence of H 2 O 2 for 6 h (A); and analysis of optical density values of p-PI3K/PI3K (B) and p-AKT/AKT (C); expression of p-PI3K, PI3K, p-AKT, and AKT in NIH/3T3 cells treated with PC capsules for 24 h and presence/absence of H 2 O 2 for 6 h (D); and analysis of optical density values of p-PI3K/PI3K (E) and p-AKT/AKT (F). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: #p < 0.05, ##p < 0.01, ###p < 0.001 (vs control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs PC group or vs H 2 O 2 group).

Techniques: Activation Assay, Expressing, Capsules, Standard Deviation, Control

PC capsules improve HUVEC and NIH/3T3 cell dysfunction by regulating PI3K/AKT signaling. HUVECs and NIH/3T3 cells were treated with PC capsules and LY294002 for 24 h and H 2 O 2 for 6 h. (A) Images of HUVEC migration exposure to H 2 O 2 , LY294002, PC capsules after 24 h; (B) quantification of percentage wound area remaining for HUVECs; (C) images of NIH/3T3 cell migration exposure to H 2 O 2 , LY294002, PC capsules after 24 h; (D) quantification of percentage wound area remaining for NIH/3T3 cells; (E–F) digital images of endothelial cell microtubule formation after treatment with H 2 O 2 , LY294002, and PC capsules. Quantification of (G) number of branch sites, (H) total lengths and (I) numbers of tube nodes (J); and analysis of optical density values of p-PI3K/PI3K (K) and p-AKT/AKT (L); expression of p-PI3K, PI3K, p-AKT, and AKT in NIH/3T3 cells treated with PC capsules, and LY294002 for 24 h and presence/absence of H 2 O 2 for 6 h (M); and analysis of optical density values of p-PI3K/PI3K (N) and p-AKT/AKT (O). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs PC group or vs LY294002 group).

Journal: Materials Today Bio

Article Title: Procyanidin capsules attenuate PI3K/AKT-mediated mitochondrial dysfunction and accelerate skin wound healing in diabetic mice

doi: 10.1016/j.mtbio.2026.103029

Figure Lengend Snippet: PC capsules improve HUVEC and NIH/3T3 cell dysfunction by regulating PI3K/AKT signaling. HUVECs and NIH/3T3 cells were treated with PC capsules and LY294002 for 24 h and H 2 O 2 for 6 h. (A) Images of HUVEC migration exposure to H 2 O 2 , LY294002, PC capsules after 24 h; (B) quantification of percentage wound area remaining for HUVECs; (C) images of NIH/3T3 cell migration exposure to H 2 O 2 , LY294002, PC capsules after 24 h; (D) quantification of percentage wound area remaining for NIH/3T3 cells; (E–F) digital images of endothelial cell microtubule formation after treatment with H 2 O 2 , LY294002, and PC capsules. Quantification of (G) number of branch sites, (H) total lengths and (I) numbers of tube nodes (J); and analysis of optical density values of p-PI3K/PI3K (K) and p-AKT/AKT (L); expression of p-PI3K, PI3K, p-AKT, and AKT in NIH/3T3 cells treated with PC capsules, and LY294002 for 24 h and presence/absence of H 2 O 2 for 6 h (M); and analysis of optical density values of p-PI3K/PI3K (N) and p-AKT/AKT (O). Data are presented as mean ± standard deviation (SD) with n = 3 independent biological replicates per group. Statistical analysis was performed using one-way ANOVA followed by Tukey's post-hoc test. Significance markers are defined as: ##p < 0.01, ###p < 0.001 (vs. control group); ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (vs PC group or vs LY294002 group).

Techniques: Capsules, Migration, Expressing, Standard Deviation, Control

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.

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

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).

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).

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

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).

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).

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

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

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).

Techniques: Cell Culture, Control, Isolation, Incubation

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.

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

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).

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).

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

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