human adult dermal microvascular endothelial cells  (ATCC)

Journal: Science Advances

Article Title: 3D bioprinting of collagen-based high-resolution internally perfusable scaffolds for engineering fully biologic tissue systems

doi: 10.1126/sciadv.adu5905

Figure Lengend Snippet: ( A ) Schematic and 3D printer machine pathing (G-code) of dual parallel channel multi-material CHIPS with an internal fibrin-based vascular cell bioink (HUVEC and MSC) region containing fluidic channels. ( B ) 3D confocal fluorescence imaging of optically cleared high-fidelity collagen (magenta) and vascular cellular bioink (green) printing with XY and XZ plane views revealing internal structure, open channels, and volumetric registration between the collagen and vascular bioink. ( C ) Schematic design and G-code of dual-channel CHIPS with collagen-reinforced walls between the cellular channels. ( D ) XY mid-plane view of whole-mount confocal fluorescence images of optically cleared 1-day statically cultured vascular CHIPS with collagen walls. Combined multiphoton fluorescence and second harmonic imaging of the vascular bioink and collagen shows spatial alignment between the cellular biomaterial (green) and collagen walls (gray). ( E ) Both XY and XZ midplane slice views from 3D confocal imaging of optically cleared dual parallel channel cellular CHIPS with fibrin walls statically cultured for 8 days and stained for actin (green), nuclei (blue), and the endothelial marker VE-Cad (red). ( F and G ) Confocal images of regions within (E) revealing microvascular cell spreading and migration (F) and vascular-like network formation (G) with inset displaying magnified views of VE-Cad only channel. ( H ) Both XY and XZ slice views of the channel bottom surface from 3D confocal imaging of optically cleared dual parallel channel cellular CHIPS with collagen walls following 8 days of perfusion culture within VAPOR. ( I and J ) Confocal images revealing strong luminal expression of VE-Cad (I) and evidence of cellular remodeling and vascular-like network maturation (J) with inset images displaying magnified views of VE-Cad and Actin only channels. *Indicates air bubble artifacts introduced during CHIPS optical clearing and imaging.

Article Snippet: Adult human dermal microvascular endothelial cells (HDMECs) (Lonza, CC-2543) were cultured on flasks coated with quick-coating solution in microvascular endothelial cell media (Lonza, EGM-2 MV).

Techniques: Fluorescence, Imaging, Cell Culture, Staining, Marker, Migration, Expressing

Journal: Angiogenesis

Article Title: Clioquinol inhibits angiogenesis by promoting VEGFR2 degradation and synergizes with AKT inhibition to suppress triple-negative breast cancer vascularization

doi: 10.1007/s10456-024-09965-1

Figure Lengend Snippet: Clioquinol selectively targets ECs and inhibits angiogenesis. a Viability (% of 0 µM) of HUVECs, HDMECs, hPC-PLs, NHDFs, MDA-MB-231, MCF-7, and 4T1-Luc2 cells after 48-hour exposure to a serial dilution of clioquinol, as assessed by WST-1 assay ( n = 4). b Cytotoxicity (% of total cell death) of clioquinol against HUVECs after 24-hour treatment, as assessed by LDH assay ( n = 4). c Proliferation (% of 0 µM) of HUVECs treated with 0, 2.5, 5, or 10 µM clioquinol for 24 h, as assessed by BrdU incorporation assay ( n = 4). d Light microscopic images of migrated HUVECs after 5-hour incubation. The cells were treated with 0, 2.5, 5, or 10 µM clioquinol for 24 h prior to this assay. Scale bar: 65 μm. e Migration (% of 0 µM) of HUVECs treated as described in (d) ( n = 3). f Phase-contrast microscopic images of tube-forming HUVECs after 18-hour treatment with 0, 2.5, 5, or 10 µM clioquinol. Scale bar: 700 μm. g Tube formation (% of 0 µM) of HUVECs treated as described in (f) ( n = 3). h Phase-contrast microscopic images of HUVEC spheroids after 24-hour treatment with 0, 2.5, 5, or 10 µM clioquinol. Scale bar: 95 μm. i Sprouting (% of 0 µM) of HUVEC spheroids treated as described in (h) ( n = 11–13). j Fluorescence microscopic images of Matrigel plugs containing DMSO (control) or 10 µM clioquinol. The sections were stained with an anti-CD31 antibody (red) and Hoechst 33342 (blue) for the visualization of ECs and cell nuclei, respectively. Scale bar: 45 μm. k Microvessel density (% of control) of control and clioquinol-containing Matrigel plugs, as assessed by immunohistochemistry ( n = 7). Means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. (a-c, e, g, i: one-way ANOVA with Tukey’s multiple comparisons test; k: unpaired Student’s t-test)

Article Snippet: Human umbilical vein endothelial cells (HUVECs), human dermal microvascular endothelial cells (HDMECs), and human pericytes from placenta (hPC-PLs) were purchased from PromoCell (Heidelberg, Germany).

Techniques: Serial Dilution, WST-1 Assay, Lactate Dehydrogenase Assay, BrdU Incorporation Assay, Incubation, Migration, Fluorescence, Control, Staining, Immunohistochemistry

Journal: Angiogenesis

Article Title: Clioquinol inhibits angiogenesis by promoting VEGFR2 degradation and synergizes with AKT inhibition to suppress triple-negative breast cancer vascularization

doi: 10.1007/s10456-024-09965-1

Figure Lengend Snippet: Clioquinol selectively down-regulates VEGFR2 in ECs. a Western blots showing VEGFR2, VEGFR1, Tie2, FGFR1, and β-actin expression in HUVECs after 4-hour treatment with 0, 2.5, 5, or 10 µM clioquinol. b - e Expression level (% of 0 µM) of VEGFR2 ( b ), VEGFR1 ( c ), Tie2 ( d ), and FGFR1 ( e ) normalized to β-actin in HUVECs treated as described in (a) ( n = 3 independent experiments). f Mean fluorescence intensity (MFI) of membrane VEGFR2 on HUVECs treated with or without 10 µM clioquinol for 0.5, 1, 2, 3, and 4 h, as assessed by flow cytometry ( n = 4). g Phase-contrast microscopic images of HUVEC spheroids after 24-hour treatment without or with clioquinol in the absence or presence of 25 ng/mL VEGF. Scale bar: 135 μm. h Sprouting (% of control) of HUVEC spheroids treated as described in (g) ( n = 13–15). i Western blots showing VEGFR2 and β-actin expression in HUVECs, HDMECs, hPC-PLs, NHDFs, MCF-7, MDA-MB-231, and 4T1-Luc2 cells. j Expression level (% of HUVEC) of VEGFR2 normalized to β-actin in different cell types as described in (i) ( n = 3 independent experiments). k Correlation between cell viability and VEGFR2 expression following exposure to 10 or 25 µM clioquinol. Means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. (b-e, h, j: one-way ANOVA with Tukey’s multiple comparisons test; f: unpaired Student’s t-test; k: Pearson correlation coefficient)

Article Snippet: Human umbilical vein endothelial cells (HUVECs), human dermal microvascular endothelial cells (HDMECs), and human pericytes from placenta (hPC-PLs) were purchased from PromoCell (Heidelberg, Germany).

Techniques: Western Blot, Expressing, Fluorescence, Membrane, Flow Cytometry, Control