human dermal microvascular endothelial cells  (PromoCell)

Journal: Angiogenesis

Article Title: Exploiting porphyrin metabolism to inhibit angiogenesis

doi: 10.1007/s10456-026-10034-y

Figure Lengend Snippet: ALA promotes porphyrin overdrive in human microvascular EC a Intracellular porphyrins levels in HMEC treated with 5 mM ALA for 4, 24, and 72 h. b – c Fluorescence imaging and relative quantification of intracellular porphyrins (magenta) in HMEC treated with 5 mM ALA and controls. Scale Bar: 50 µm. n = 12 d Intracellular heme levels in HMEC treated with 5 mM ALA for 4, 24, and 72 h. e Mitochondrial activity of ALAS in HMEC treated with 5 mM ALA for 24 h (hrs). f – h Representative Western Blot images ( f ) and their quantification g – h of ALAS1 and HO-1 protein levels in HMEC treated with 5 mM ALA for 24 and 72 h. i – j Differential expression of heme-related genes in HMEC upon 24 h of 5 mM ALA treatment. The heatmap shown in ( i ) displays differences in gene expression levels, represented as fold change (FC) respect to not-treated (NT) cells. Volcano plot shown in ( j ) represents the differences in gene expression levels versus -log10(q value). The dotted line indicates the significance threshold of FDR q < 1% (-log10(q value) = 2). The plotted results represent mean values obtained from at least 5 individual biological replicates. k – l Extracellular levels of porphyrins ( k ) and heme ( l ) in HMEC after 4, 24, and 72 h of 5 mM ALA supplementation. Data are expressed as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For statistical analyses, ordinary one-way ANOVA test with Tukey’s multiple comparisons ( a , d , g , h , k , l ) and parametric unpaired t test ( c , e ) were used. FC, fold change; NT, not treated controls; hrs, hours; ALA, 5-amminolevulinic acid

Article Snippet: Human adult dermal microvascular endothelial cells (HMEC-1, RRID:CVCL_0307) were purchased by ATCC, propagated in MCDB131 (Thermo Fisher Scientific, Waltham, MA USA, catalog n°10,372,019) supplemented with 10% heat-inactivated low-endotoxin FBS (GIBCO by Thermofisher Scientific, Waltham, MA USA, catalog n10270106), 10 mM GlutaMAXTM Supplement (Thermo Fisher Scientific, Waltham, MA USA, catalog n°35,050,061), 10 ng/mL Epidermal Growth Factor (Thermo Fisher Scientific, Waltham, MA USA, catalog n° PHG0315), 1 μg/mL Hydrocortisone-Water Soluble (Sigma Aldrich, St. Louis, MO USA, catalog n° H0396) and used up to passage 12.

Techniques: Fluorescence, Imaging, Quantitative Proteomics, Activity Assay, Western Blot, Gene Expression

Journal: Angiogenesis

Article Title: Exploiting porphyrin metabolism to inhibit angiogenesis

doi: 10.1007/s10456-026-10034-y

Figure Lengend Snippet: ALA inhibits in vitro angiogenesis on human microvascular EC ( a and b ) Proliferation of HMEC treated with increasing concentrations of ALA (500 nM, 0.1 mM, 5 mM) at various time points. Scale bar: 500 µm. n = 6 (c and d) Wound healing experiment of HMEC treated with increasing concentrations of ALA (500 nM, 0.1 mM, 5 mM) at various time points. Scale bar: 500 µm. n > 8 (e – h) in vitro tubulogenesis assay performed on 5 mM ALA-treated and control HMEC. Quantification of the total length ( f ) of the networks, number of nodes ( g ) and number of branches ( h ) are shown. Scale bar: 500 µm. Data are expressed as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For statistical analyses ordinary two-way ANOVA test with Tukey’s multiple comparisons ( b , d ) and parametric unpaired t-test ( f – h ) were used. FC, fold change; NT, not treated controls; hrs, hours; ALA, 5-amminolevulinic acid

Article Snippet: Human adult dermal microvascular endothelial cells (HMEC-1, RRID:CVCL_0307) were purchased by ATCC, propagated in MCDB131 (Thermo Fisher Scientific, Waltham, MA USA, catalog n°10,372,019) supplemented with 10% heat-inactivated low-endotoxin FBS (GIBCO by Thermofisher Scientific, Waltham, MA USA, catalog n10270106), 10 mM GlutaMAXTM Supplement (Thermo Fisher Scientific, Waltham, MA USA, catalog n°35,050,061), 10 ng/mL Epidermal Growth Factor (Thermo Fisher Scientific, Waltham, MA USA, catalog n° PHG0315), 1 μg/mL Hydrocortisone-Water Soluble (Sigma Aldrich, St. Louis, MO USA, catalog n° H0396) and used up to passage 12.

Techniques: In Vitro, Control

Journal: bioRxiv

Article Title: Light-driven repair: Photobiomodulation restores blood–brain barrier function following hypoxic injury

doi: 10.64898/2026.02.15.706027

Figure Lengend Snippet: A) Experimental workflow illustrating the transwell-based BBB model, hypoxic exposure (6 h, 1% O₂), PBM treatment schedule, and downstream functional and molecular analyses (created with BioRender). (B–E) TEER expressed as relative change (%) from baseline. (B) Normoxic controls at 48 h. (C)TEER immediately following hypoxia. (D)TEER at 24 h post-hypoxia. (E)TEER at 48 h post-hypoxia. PBM significantly restored endothelial barrier resistance under hypoxic conditions. Statistical comparisons were performed using multiple unpaired t-tests with Holm–Šídák correction; N=4 independent biological replicates; *p<0.05. (F)ZO-1 mRNA expression in HBMECs at 48 h, normalised to RPL13A. (G)Quantification of ZO-1 protein levels relative to control, measured as ZO-1-positive area normalised to Hoechst nuclear area. (H)Representative ICC images showing ZO-1 (magenta) and nuclei (Hoechst, blue) in normoxic and hypoxic HBMECs with and without PBM. For mRNA and protein analyses, statistical significance was determined using one-way ANOVA with Šídák’s post hoc correction; N=3–4 independent biological replicates. Data are presented as mean±SEM.

Article Snippet: The cell lines used were purchased from Innoprot: human brain microvascular endothelial cells (HBMECs) (Innoprot; P10361-IM), human astrocytes (HAs) (Innoprot; P10251-IM), and human brain vascular pericytes (HVPCs) (Innoprot; P10363-IM).

Techniques: Functional Assay, Expressing, Control

Journal: bioRxiv

Article Title: Light-driven repair: Photobiomodulation restores blood–brain barrier function following hypoxic injury

doi: 10.64898/2026.02.15.706027

Figure Lengend Snippet: (A)vWF mRNA expression in HBMECs 48 h after normoxia or hypoxia (6 h, 1% O₂), with or without PBM treatment, normalised to RPL13A. (B)Representative ICC images showing vWF (yellow) and nuclei (Hoechst, blue) in normoxic and hypoxic endothelial cells ±PBM. (C)Quantification of vWF protein levels, expressed as mean nuclear-normalised fluorescence intensity relative to normoxia −PBM controls. (D)Validation of vWF knockdown efficiency in siRNA-transfected endothelial cells, shown as relative vWF mRNA expression normalised to RPL13A. (E-F) Relative TEER changes (%) in endothelial monocultures and BBB tri-cultures at 24 h and 48 h under normoxic (E) and hypoxic (F) conditions following vWF silencing. Statistical comparisons for mRNA and protein expression were performed using one-way ANOVA with Šidák’s post hoc test (N=3–4 biological replicates). siRNA validation was analysed using an unpaired two-tailed t-test (N=8 biological replicates). TEER data were analysed using multiple unpaired t-tests with Holm–Šidák correction (N=4 biological replicates). Data are presented as mean±SEM.

Article Snippet: The cell lines used were purchased from Innoprot: human brain microvascular endothelial cells (HBMECs) (Innoprot; P10361-IM), human astrocytes (HAs) (Innoprot; P10251-IM), and human brain vascular pericytes (HVPCs) (Innoprot; P10363-IM).

Techniques: Expressing, Fluorescence, Biomarker Discovery, Knockdown, Transfection, Two Tailed Test