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primary human dermal microvascular endothelial cells hmvec d  (Lonza)


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    Lonza primary human dermal microvascular endothelial cells hmvec d
    Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human <t>microvascular</t> <t>endothelial</t> cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.
    Primary Human Dermal Microvascular Endothelial Cells Hmvec D, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Transendothelial migration of the Lyme disease spirochete involves spirochete internalization as an intermediate step through a transcellular pathway that involves Cdc42 and Rac1"

    Article Title: Transendothelial migration of the Lyme disease spirochete involves spirochete internalization as an intermediate step through a transcellular pathway that involves Cdc42 and Rac1

    Journal: Microbiology Spectrum

    doi: 10.1128/spectrum.02221-24

    Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human microvascular endothelial cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.
    Figure Legend Snippet: Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human microvascular endothelial cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.

    Techniques Used: Migration, Transmigration Assay, Diffusion-based Assay, Infection, Flow Cytometry, MANN-WHITNEY, Microscopy, Control, Comparison



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    Lonza primary human dermal microvascular endothelial cells hmvec d
    Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human <t>microvascular</t> <t>endothelial</t> cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.
    Primary Human Dermal Microvascular Endothelial Cells Hmvec D, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MTS assay of HaCaT keratinocytes ( A – C ), human dermal fibroblasts (HDF) ( D – F ), and human dermal <t>microvascular</t> vein <t>endothelial</t> cells (HMVEC-d) ( G – I ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d. The figures were generated using GraphPad Prism software (GraphPad Software, San Diego, CA, USA), which was also utilized to logarithmically transform the concentration values and generate fitting curves to visualize the data.
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    MTS assay of HaCaT keratinocytes ( A – C ), human dermal fibroblasts (HDF) ( D – F ), and human dermal <t>microvascular</t> vein <t>endothelial</t> cells (HMVEC-d) ( G – I ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d. The figures were generated using GraphPad Prism software (GraphPad Software, San Diego, CA, USA), which was also utilized to logarithmically transform the concentration values and generate fitting curves to visualize the data.
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    MTS assay of HaCaT keratinocytes ( A – C ), human dermal fibroblasts (HDF) ( D – F ), and human dermal <t>microvascular</t> vein <t>endothelial</t> cells (HMVEC-d) ( G – I ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d. The figures were generated using GraphPad Prism software (GraphPad Software, San Diego, CA, USA), which was also utilized to logarithmically transform the concentration values and generate fitting curves to visualize the data.
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    <t>HMVEC-d</t> proliferation response was greater when exposed to EGLN1 KO (Sg1) HEK-293T preconditioned media compared to exposure to unmodified (Neg) HEK-293T preconditioned media in both normoxic and hypoxic conditions. Columns represent mean, points represent individual wells (n=6), bars represent standard deviation. * represents significance at p<0.05.
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    TaKaRa human dermal microvascular endothelial cells hmvec d
    Effects of deoxyshikonin on cord formation of HMVEC-dLy and <t>HMVEC-d</t> on Matrigel. (a), (c) Photographs of cord formation of HMVEC-dLy and HMVEC-d on Matrigel after incubation with or without 0.8 μ M deoxyshikonin at 2 to 6 h (at ×400 magnification). (b), (d) The relative length of cords was measured using an Angiogenesis Image Analyzer. Data are the mean ± SD ( n = 3); * P < 0.05, ** P < 0.01 compared with the control.
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    The effects of MEK inhibitors on SSRBC adhesion to HUVECs, <t>HMVECs-d</t> and EOMA cells was tested in intermittent flow condition assays at different shear stresses in vitro . Results are presented as % adherent SSRBCs at a shear stress of 2 dynes/cm 2 . A. SSRBCs were sham-treated or treated with 100 nM MEK inhibitor U0126 prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; ***: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs. Error bars show standard error mean (SEM) of 4 different experiments. B. SSRBCs were sham-treated, or treated with 100 nM RDEA119, 100 nM AZD6244, 100 nM trametinib, or 10 µM damnacanthal prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs; and † : p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs. Error bars show SEM of 3 different experiments. C – D. SSRBCs and normal RBCs (AARBCs) were sham-treated, or treated with 100 nM U0126 or 100 nM RDEA119 prior to adhesion assays to non-treated and TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). *: p <0.0001 compared to sham-treated AARBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); **: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); and ***: p <0.001 compared to sham-treated SSRBCs adherent to TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). Error bars show SEM of 3 different experiments for C and D .
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    Lonza dermal human microvascular endothelial cells hmvec d
    (A) <t>HMVEC-D</t> were cultured on collagen I and permeability was assessed using a Transwell assay (N = 3/group). Endothelial cells were serum starved (1 hr) and FITC labeled albumin was added to the top well and cells were incubated. QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to induce permeability and the amount of FITC-linked albumin in the bottom well was quantified (Pa/hr) (3 hr). Peptide QHREDGS reduced HMVEC-D permeability (*P = 0.004) compared to control and scrambled peptide (**P = 0.001) groups. (B) Transendothelial electrical resistance was measured in HMVEC-D cultured on collagen I. HMVEC-D were serum starved and QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to reduce resistance and resistance was measured as a function of time (N = 2/group). Peptide QHREDGS increased QHREDGS resistance compared to control and scrambled peptides. Red coloured triangles indicate normalized resistance values for the QHREDGS treated group.
    Dermal Human Microvascular Endothelial Cells Hmvec D, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human microvascular endothelial cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.

    Journal: Microbiology Spectrum

    Article Title: Transendothelial migration of the Lyme disease spirochete involves spirochete internalization as an intermediate step through a transcellular pathway that involves Cdc42 and Rac1

    doi: 10.1128/spectrum.02221-24

    Figure Lengend Snippet: Transendothelial migration of B. burgdorferi in Transwell chambers. (A) To study B. burgdorferi transmigration, Transwell chambers were seeded with hMVEC-d or hTERT cells until a tight monolayer was formed (<4% albumin diffusion). The upper chamber was infected with 3 × 10 5 spirochetes; after 20 h of infection, we counted the spirochetes in both the upper and lower chambers (either manually or by flow cytometry) and calculated the percentage of total transmigrated spirochetes (lower chamber). ( B, C ) The graphs show the percentage (mean ± SD) of B. burgdorferi that had transmigrated through human microvascular endothelial cells as determined by counting spirochetes in both the upper and lower chamber by flow cytometry. The data represent the mean of % transmigration ±SD of three independent experiments performed in quadruplicate and analyzed for significance using the Mann–Whitney test. NI denotes the non-infectious strain GCB705 (B31-A + pTM61 gent,gfp, see ), and WT indicates GCB726. ( D ) Evaluation of B. burgdorferi transendothelial migration in hTERT treated with AKB-9785. The complete chamber was treated with 5 µM AKB9785 for the duration of the assay. The upper chamber was infected with 3 × 10 5 spirochetes, and the percentage of total transmigrated spirochete was assessed by counting in a Petroff–Hausser chamber and dark-field microscopy after 20 h of infection. The data represent the mean % transmigration ±SD of three independent experiments performed in quadruplicate; non-parametric ANOVA was performed to compare the control cells with the treated cells using the Kruskal–Wallis post-test (ns = not significant). ( E ) Demonstration of the effectiveness of AKB-9785 to lock intercellular junctions in hTERT cells. Ly 294002 at 40 µM was used to disrupt the monolayer, and AKB-9785 was used to lock intercellular junctions and preserve monolayer integrity, which was assessed using an albumin diffusion assay: 10 μg of 555-Alb was added to the upper chamber at 16 h, and the reading was carried out at the final point (20 h). The graph represents the mean ± SD of three experiments, with 1–4 samples for each experimental condition. Statistics were evaluated with the Kruskal–Wallis test and Dunn’s multiple comparison; P < 0.05 was considered significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = not significant.

    Article Snippet: Primary human dermal microvascular endothelial cells (hMVEC-d) were purchased from Lonza (CC-2543), grown in Basal Medium (EBM-2) (CC-3156, Lonza) complete media (with supplements EGMTM-2 SingleQuotsTM Supplements [CC-4176, Lonza]) at 37°C under 5% CO 2 and used before passage five. hTERT-immortalized dermal microvascular endothelial cell, neonatal (CRL4060, ATCC), was used. hTERT was cultured in Vascular Cell Basal Medium (VCBM) (PCS-100-030, ATCC) with the microvascular endothelial cell growth kit-BBE (PCS-110-040, ATCC) +0.5 μg/mL puromycin (P8833, Sigma-Aldrich) according to the manufacturer’s instruction.

    Techniques: Migration, Transmigration Assay, Diffusion-based Assay, Infection, Flow Cytometry, MANN-WHITNEY, Microscopy, Control, Comparison

    MTS assay of HaCaT keratinocytes ( A – C ), human dermal fibroblasts (HDF) ( D – F ), and human dermal microvascular vein endothelial cells (HMVEC-d) ( G – I ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d. The figures were generated using GraphPad Prism software (GraphPad Software, San Diego, CA, USA), which was also utilized to logarithmically transform the concentration values and generate fitting curves to visualize the data.

    Journal: Biomolecules

    Article Title: Assessment of Agrimonia eupatoria L. and Lipophosphonoxin (DR-6180) Combination for Wound Repair: Bridging the Gap Between Phytomedicine and Organic Chemistry

    doi: 10.3390/biom14121590

    Figure Lengend Snippet: MTS assay of HaCaT keratinocytes ( A – C ), human dermal fibroblasts (HDF) ( D – F ), and human dermal microvascular vein endothelial cells (HMVEC-d) ( G – I ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d. The figures were generated using GraphPad Prism software (GraphPad Software, San Diego, CA, USA), which was also utilized to logarithmically transform the concentration values and generate fitting curves to visualize the data.

    Article Snippet: Dermal microvascular endothelial cells (HMVEC-d) were obtained from Lonza (Lonza Walkersville, Inc., Walkersville, MD, USA).

    Techniques: MTS Assay, Positive Control, Generated, Software, Concentration Assay

    Wound healing (2D migration) assay of HaCaT keratinocytes ( A ) and human dermal microvascular vein endothelial cells—HMVEC-d ( B ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT, while VEGF-A was used in HMVEC-d (Magnification 100×).

    Journal: Biomolecules

    Article Title: Assessment of Agrimonia eupatoria L. and Lipophosphonoxin (DR-6180) Combination for Wound Repair: Bridging the Gap Between Phytomedicine and Organic Chemistry

    doi: 10.3390/biom14121590

    Figure Lengend Snippet: Wound healing (2D migration) assay of HaCaT keratinocytes ( A ) and human dermal microvascular vein endothelial cells—HMVEC-d ( B ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT, while VEGF-A was used in HMVEC-d (Magnification 100×).

    Article Snippet: Dermal microvascular endothelial cells (HMVEC-d) were obtained from Lonza (Lonza Walkersville, Inc., Walkersville, MD, USA).

    Techniques: Migration, Positive Control

    Western blot (WB) analysis of HaCaT keratinocytes ( A ), human dermal fibroblasts—HDF ( B ), and human dermal microvascular vein endothelial cells—HMVEC-d ( C ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d.

    Journal: Biomolecules

    Article Title: Assessment of Agrimonia eupatoria L. and Lipophosphonoxin (DR-6180) Combination for Wound Repair: Bridging the Gap Between Phytomedicine and Organic Chemistry

    doi: 10.3390/biom14121590

    Figure Lengend Snippet: Western blot (WB) analysis of HaCaT keratinocytes ( A ), human dermal fibroblasts—HDF ( B ), and human dermal microvascular vein endothelial cells—HMVEC-d ( C ) in the presence of Agrimonia eupatoria L. (AE) extract, lipophosphonoxin (LPPO) DR-6180, and combination of AE and LPPO. TGF-β1 was used as the positive control in HaCaT and HDF, while VEGF-A was used in HMVEC-d.

    Article Snippet: Dermal microvascular endothelial cells (HMVEC-d) were obtained from Lonza (Lonza Walkersville, Inc., Walkersville, MD, USA).

    Techniques: Western Blot, Positive Control

    HMVEC-d proliferation response was greater when exposed to EGLN1 KO (Sg1) HEK-293T preconditioned media compared to exposure to unmodified (Neg) HEK-293T preconditioned media in both normoxic and hypoxic conditions. Columns represent mean, points represent individual wells (n=6), bars represent standard deviation. * represents significance at p<0.05.

    Journal: bioRxiv

    Article Title: Harnessing EGLN1 Gene Editing to Amplify HIF-1α and Enhance Human Angiogenic Response

    doi: 10.1101/2023.05.29.542734

    Figure Lengend Snippet: HMVEC-d proliferation response was greater when exposed to EGLN1 KO (Sg1) HEK-293T preconditioned media compared to exposure to unmodified (Neg) HEK-293T preconditioned media in both normoxic and hypoxic conditions. Columns represent mean, points represent individual wells (n=6), bars represent standard deviation. * represents significance at p<0.05.

    Article Snippet: Adult human dermal microvascular endothelial cells (HMVEC-d) (LONZA, Cat# CC-2543) (passage 8) were cultured until ∼80% confluence before being passaged and seeded at 10,000 cells/cm 2 in EGM-2MV (LONZA, Cat# CC-3202) in 6-well tissue culture plates and were allowed to adhere for 5 hours before induction of serum starvation in EBM-2 (LONZA, Cat# CC-3156) for 16 hours prior to being administered media of interest, as described below.

    Techniques: Standard Deviation

    HMVEC-d scratch closure was greater when exposed to preconditioned media from EGLN1 KO (Sg1) HEK-293T cells versus unmodified (Neg) HEK-293T cells in both normoxic or hypoxic conditions. Columns represent mean, points represent individual image points (n=6-9), bars represent standard deviation.

    Journal: bioRxiv

    Article Title: Harnessing EGLN1 Gene Editing to Amplify HIF-1α and Enhance Human Angiogenic Response

    doi: 10.1101/2023.05.29.542734

    Figure Lengend Snippet: HMVEC-d scratch closure was greater when exposed to preconditioned media from EGLN1 KO (Sg1) HEK-293T cells versus unmodified (Neg) HEK-293T cells in both normoxic or hypoxic conditions. Columns represent mean, points represent individual image points (n=6-9), bars represent standard deviation.

    Article Snippet: Adult human dermal microvascular endothelial cells (HMVEC-d) (LONZA, Cat# CC-2543) (passage 8) were cultured until ∼80% confluence before being passaged and seeded at 10,000 cells/cm 2 in EGM-2MV (LONZA, Cat# CC-3202) in 6-well tissue culture plates and were allowed to adhere for 5 hours before induction of serum starvation in EBM-2 (LONZA, Cat# CC-3156) for 16 hours prior to being administered media of interest, as described below.

    Techniques: Standard Deviation

    Effects of deoxyshikonin on cord formation of HMVEC-dLy and HMVEC-d on Matrigel. (a), (c) Photographs of cord formation of HMVEC-dLy and HMVEC-d on Matrigel after incubation with or without 0.8 μ M deoxyshikonin at 2 to 6 h (at ×400 magnification). (b), (d) The relative length of cords was measured using an Angiogenesis Image Analyzer. Data are the mean ± SD ( n = 3); * P < 0.05, ** P < 0.01 compared with the control.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Enhancement of Lymphangiogenesis In Vitro via the Regulations of HIF-1 α Expression and Nuclear Translocation by Deoxyshikonin

    doi: 10.1155/2013/148297

    Figure Lengend Snippet: Effects of deoxyshikonin on cord formation of HMVEC-dLy and HMVEC-d on Matrigel. (a), (c) Photographs of cord formation of HMVEC-dLy and HMVEC-d on Matrigel after incubation with or without 0.8 μ M deoxyshikonin at 2 to 6 h (at ×400 magnification). (b), (d) The relative length of cords was measured using an Angiogenesis Image Analyzer. Data are the mean ± SD ( n = 3); * P < 0.05, ** P < 0.01 compared with the control.

    Article Snippet: Human dermal lymphatic microvascular endothelial cells (HMVEC-dLy) and human dermal microvascular endothelial cells (HMVEC-d) were obtained from Takara Bio Inc. (Shiga, Japan).

    Techniques: Incubation

    The effects of MEK inhibitors on SSRBC adhesion to HUVECs, HMVECs-d and EOMA cells was tested in intermittent flow condition assays at different shear stresses in vitro . Results are presented as % adherent SSRBCs at a shear stress of 2 dynes/cm 2 . A. SSRBCs were sham-treated or treated with 100 nM MEK inhibitor U0126 prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; ***: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs. Error bars show standard error mean (SEM) of 4 different experiments. B. SSRBCs were sham-treated, or treated with 100 nM RDEA119, 100 nM AZD6244, 100 nM trametinib, or 10 µM damnacanthal prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs; and † : p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs. Error bars show SEM of 3 different experiments. C – D. SSRBCs and normal RBCs (AARBCs) were sham-treated, or treated with 100 nM U0126 or 100 nM RDEA119 prior to adhesion assays to non-treated and TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). *: p <0.0001 compared to sham-treated AARBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); **: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); and ***: p <0.001 compared to sham-treated SSRBCs adherent to TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). Error bars show SEM of 3 different experiments for C and D .

    Journal: PLoS ONE

    Article Title: MEK Inhibitors, Novel Anti-Adhesive Molecules, Reduce Sickle Red Blood Cell Adhesion In Vitro and In Vivo, and Vasoocclusion In Vivo

    doi: 10.1371/journal.pone.0110306

    Figure Lengend Snippet: The effects of MEK inhibitors on SSRBC adhesion to HUVECs, HMVECs-d and EOMA cells was tested in intermittent flow condition assays at different shear stresses in vitro . Results are presented as % adherent SSRBCs at a shear stress of 2 dynes/cm 2 . A. SSRBCs were sham-treated or treated with 100 nM MEK inhibitor U0126 prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; ***: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs. Error bars show standard error mean (SEM) of 4 different experiments. B. SSRBCs were sham-treated, or treated with 100 nM RDEA119, 100 nM AZD6244, 100 nM trametinib, or 10 µM damnacanthal prior to adhesion assays to non-treated and TNFα-treated HUVECs. *: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HUVECs; **: p <0.0001 compared to sham-treated SSRBCs adherent to TNFα-treated HUVECs; and † : p <0.001 compared to sham-treated SSRBCs adherent to non-treated HUVECs. Error bars show SEM of 3 different experiments. C – D. SSRBCs and normal RBCs (AARBCs) were sham-treated, or treated with 100 nM U0126 or 100 nM RDEA119 prior to adhesion assays to non-treated and TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). *: p <0.0001 compared to sham-treated AARBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); **: p <0.0001 compared to sham-treated SSRBCs adherent to non-treated HMVECs-d ( C ) and EOMA cells ( D ); and ***: p <0.001 compared to sham-treated SSRBCs adherent to TNFα-treated HMVECs-d ( C ) and EOMA cells ( D ). Error bars show SEM of 3 different experiments for C and D .

    Article Snippet: Human umbilical vein endothelial cells (HUVECs, ATCC), the murine endothelial cell line EOMA (ATCC, Manassas, VA), which exhibits properties characteristic of microvascular endothelial cells, and human dermal microvascular endothelial cells (HMVECs-d) (Lonza, Walkersville, MD) were grown as monolayers in EBM2 medium (Clonetics, Walkersville, MD) supplemented with EGM2 (Clonetics) , .

    Techniques: In Vitro

    The effect of MEK inhibitors on the ability of SSRBCs to stimulate neutrophil (PMN) adhesion to ECs was tested. A and B . SSRBCs (n = 8) were sham-treated or treated with 100 nM MEK inhibitor U0126, RDEA119, AZD6244 or trametinib. Washed treated SSRBCs were then co-incubated with ABO-matched naïve PMNs isolated from healthy donors (n = 8), prior to testing adhesion of PMNs to HUVECs ( A; n = 4) and HMVECs-d ( B; n = 4) in intermittent flow condition assays at different shear stresses. C. AARBCs (n = 3) were sham-treated, washed, and then co-incubated with ABO-matched naïve PMNs isolated from healthy donors (n = 3), prior to testing adhesion of PMNs to HUVECs and HMVECs-d at different shear stresses. D. Non-treated and TNFα-treated HUVECs were co-incubated with sham-treated SSRBCs, U0126-treated SSRBCs or sham-treated AARBCs. HUVECs were then washed free of non-adherent RBCs, and tested for their ability to support adhesion of PMNs (n = 3). Results are presented as % adherent PMNs at a shear stress of 1 dyne/cm 2 . *: p <0.0001 compared to adhesion of naïve PMNs (PMNs only) to non-treated ECs; and **: p <0.0001 compared to adhesion of PMNs stimulated with SSRBCs (PMNs+SSRBCs). Error bars show SEM of 4 different experiments for A and B , and 3 different experiments for C and D .

    Journal: PLoS ONE

    Article Title: MEK Inhibitors, Novel Anti-Adhesive Molecules, Reduce Sickle Red Blood Cell Adhesion In Vitro and In Vivo, and Vasoocclusion In Vivo

    doi: 10.1371/journal.pone.0110306

    Figure Lengend Snippet: The effect of MEK inhibitors on the ability of SSRBCs to stimulate neutrophil (PMN) adhesion to ECs was tested. A and B . SSRBCs (n = 8) were sham-treated or treated with 100 nM MEK inhibitor U0126, RDEA119, AZD6244 or trametinib. Washed treated SSRBCs were then co-incubated with ABO-matched naïve PMNs isolated from healthy donors (n = 8), prior to testing adhesion of PMNs to HUVECs ( A; n = 4) and HMVECs-d ( B; n = 4) in intermittent flow condition assays at different shear stresses. C. AARBCs (n = 3) were sham-treated, washed, and then co-incubated with ABO-matched naïve PMNs isolated from healthy donors (n = 3), prior to testing adhesion of PMNs to HUVECs and HMVECs-d at different shear stresses. D. Non-treated and TNFα-treated HUVECs were co-incubated with sham-treated SSRBCs, U0126-treated SSRBCs or sham-treated AARBCs. HUVECs were then washed free of non-adherent RBCs, and tested for their ability to support adhesion of PMNs (n = 3). Results are presented as % adherent PMNs at a shear stress of 1 dyne/cm 2 . *: p <0.0001 compared to adhesion of naïve PMNs (PMNs only) to non-treated ECs; and **: p <0.0001 compared to adhesion of PMNs stimulated with SSRBCs (PMNs+SSRBCs). Error bars show SEM of 4 different experiments for A and B , and 3 different experiments for C and D .

    Article Snippet: Human umbilical vein endothelial cells (HUVECs, ATCC), the murine endothelial cell line EOMA (ATCC, Manassas, VA), which exhibits properties characteristic of microvascular endothelial cells, and human dermal microvascular endothelial cells (HMVECs-d) (Lonza, Walkersville, MD) were grown as monolayers in EBM2 medium (Clonetics, Walkersville, MD) supplemented with EGM2 (Clonetics) , .

    Techniques: Incubation, Isolation

    (A) HMVEC-D were cultured on collagen I and permeability was assessed using a Transwell assay (N = 3/group). Endothelial cells were serum starved (1 hr) and FITC labeled albumin was added to the top well and cells were incubated. QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to induce permeability and the amount of FITC-linked albumin in the bottom well was quantified (Pa/hr) (3 hr). Peptide QHREDGS reduced HMVEC-D permeability (*P = 0.004) compared to control and scrambled peptide (**P = 0.001) groups. (B) Transendothelial electrical resistance was measured in HMVEC-D cultured on collagen I. HMVEC-D were serum starved and QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to reduce resistance and resistance was measured as a function of time (N = 2/group). Peptide QHREDGS increased QHREDGS resistance compared to control and scrambled peptides. Red coloured triangles indicate normalized resistance values for the QHREDGS treated group.

    Journal: PLoS ONE

    Article Title: QHREDGS Enhances Tube Formation, Metabolism and Survival of Endothelial Cells in Collagen-Chitosan Hydrogels

    doi: 10.1371/journal.pone.0072956

    Figure Lengend Snippet: (A) HMVEC-D were cultured on collagen I and permeability was assessed using a Transwell assay (N = 3/group). Endothelial cells were serum starved (1 hr) and FITC labeled albumin was added to the top well and cells were incubated. QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to induce permeability and the amount of FITC-linked albumin in the bottom well was quantified (Pa/hr) (3 hr). Peptide QHREDGS reduced HMVEC-D permeability (*P = 0.004) compared to control and scrambled peptide (**P = 0.001) groups. (B) Transendothelial electrical resistance was measured in HMVEC-D cultured on collagen I. HMVEC-D were serum starved and QHREDGS or scrambled (DQSHER) peptides at 400 µM or control (PBS) were added (1 hr). Thrombin was added to reduce resistance and resistance was measured as a function of time (N = 2/group). Peptide QHREDGS increased QHREDGS resistance compared to control and scrambled peptides. Red coloured triangles indicate normalized resistance values for the QHREDGS treated group.

    Article Snippet: Dermal human microvascular endothelial cells (HMVEC-D) were obtained (Lonza) and cultured as recommended in the manufacturer’s instructions in full endothelial growth media.

    Techniques: Cell Culture, Permeability, Transwell Assay, Labeling, Incubation