Journal: Advanced Science

Article Title: Subventricular Zone‐on‐a‐Chip: A Model to Study Neurogenesis Disruption in Neonatal Intraventricular Hemorrhage

doi: 10.1002/advs.202502145

Figure Lengend Snippet: Overview of the SVZ‐on‐a‐chip. A) The SVZ‐on‐a‐chip structure 24 h post‐seeding, showing distinct cellular compartments visualized via staining. The entire chip structure is shown (scale bar = 3000 µm) alongside a magnified maximum Z‐projection view (scale bar = 100 µm). Arrows indicate neural stem cells (NSCs), and triangles highlight human fetal astrocytes (HFAs) embedded in Matrigel, occupying the central channel. The right channel is lined with human brain microvascular endothelial cells (HBMECs), while the left channel contains human choroid plexus epithelial cells (HCPEpiC). B) Neural channel with HCPEpiC seeded along the central channel (scale bar = 200 µm). C) The central channel, highlighted by triangles, is shown in closer detail (scale bar = 200 µm). D) 3D view of the inner channel, showing cells stained with Nestin (gray). Astrocytes, marked by triangles, are stained red but lack Nestin expression (scale bar = 70 µm). E) Inner channel viewed from a different angle (scale bar = 70 µm). F) Vascular channel lined with HBMECs (scale bar = 70 µm). G) HBMECs fully covering the vascular channel (scale bar = 70 µm). H) A magnified maximum Z‐projection view of the SVZ‐on‐a‐chip 7 days after complete seeding, showing significant cellular growth (scale bar = 100 µm). Clusters of NSCs are observed invading both the neural and, predominantly, the vascular channel. I) Maximum Z‐projection view of Nestin staining in the middle channel (scale bar = 100 µm). J) Close‐up of DCX‐positive neural stem cells invading the vascular channel. DCX staining was used instead of Nestin because HBMECs also express Nestin (scale bar = 50 µm). SVZ: Subventricular Zone, NSCs: Neural Stem Cells, HFAs: Human Fetal Astrocytes, HBMECs: Human Brain Microvascular Endothelial Cells, HCPEpiC: Human Choroid Plexus Epithelial Cells, DCX: Doublecortin.

Article Snippet: DPBS minus Ca ++ and M g++ (14 190 144, Thermo Fischer, Waltham, Massachusetts, USA), DPBS with Ca ++ and Mg ++ (14 040 091, Thermo Fischer), Astrocyte Media (AM1801, ScienCell, Carlsbad, California, USA), TrypLE (12 604 013, Thermo Fischer), Attachment factor protein 1× (S006100, Thermo Fischer), EGM ‐2 MV Microvascular Endothelial Cell Growth Medium‐2 BulletKit (CC‐3202, Lonza, Basel, Switzerland), Endothelial Cell Growth Medium MV2 (C‐22022, PromoCell, Heidelberg, Germany), Epithelial Cell Medium (Innoprot, P60104 , Derio, Spain), poly‐L‐lysine (PLL) (Innoprot), DMEM: F12 Glutamax (31331‐028, Thermo Fischer), N2 supplement ( 17502‐048, Thermo Fischer), B27 serum free (11 530 536, Thermo Fischer), FGF (233‐FB, R&D Systems, MN, USA), EGF (E9644, Sigma Aldrich, Burlington, Massachusetts, USA), PLO (P3655, Sigma Aldrich), Laminin (L2020, Sigma Aldrich), Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix, LDEV‐free (354 230, Corning, Corning, New York, USA), Antibiotic‐Antimycotic (15 240 062, Themo Fischer), idenTx 3 Chip (Aim Biotech, Singapore), Transwells 0.4 μM microporous membrane (833 932 041, Sarstedt, Nümbrecht, Germany), High Pure RNA Isolation Kit (11 828 665 001, Roche, Basel, Switzerland), Clariom S Affymetrix Assay (902 927, Thermo Fischer), High‐capacity RNA‐to‐cDNA kit (4 387 406, Thermo Fischer), TaqMan probes (Applied Biosystems, California, USA), Fast Advanced Master Mix (4 444 557, Applied Biosystems), TRIzol (15 596 026, Thermo Fischer), Chloroform (194 002, MP Biomedicals, Santa Ana, California, USA), Cell recovery solution (354 253, Corning), Anti‐Adherence Rinsing Solution (0 7010, STEMCELL Technologies, Vancouver, Canada), AggreWell800 24‐well plates (34 811, STEMCELL Technologies), Goat serum (G9023, Sigma Aldrich), Trypan Blue (1 450 021, Bio‐Rad, Hercules, California, USA), LEGENDplex Human Inflammation Panel 1 (13‐plex) assay using a V‐bottom plate (BioLegend, San Diego, California, USA), CellROX Green reagent ( C10444 ,Invitrogen), JC‐1 dye (T3168, Invitrogen) IL1receptor antagonist (SRP3327, Sigma Aldrich).

Techniques: Staining, Expressing

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: (A, B) Brightfield images of cell alignment analysis. 0° and 180° are parallel to the direction of flow while 90° is perpendicular to the direction of flow. (C–F) Histograms of cell alignment under static, 1 dyne/cm 2 , 4 dyne/cm 2 , and 10 dyne/cm 2 conditions with angles grouped in cohorts every 30°. Statistical analysis was performed via analysis of variance with multiple condition correction. HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell. ∗ P ≤ .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques:

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: (A) RNA-seq PCA of all EC types. Dynes are demarcated by different colors and EC type by different shapes. (B) Volcano plot of differentially expressed genes. Log 2 fold change (FC) is expressed as the ratio of expression of shear stress samples relative to control samples. The horizontal dashed line demarcates P = 10e-20, and the vertical dashed line represents a FC cutoff of 1.5 or −1.5. (C) Venn diagram demonstrating DEGs when comparing shear stress vs static conditions in 3 EC types, HUVECs are shown in green, HPAECs in orange, and HMVECs in purple. (D) Venn diagram demonstrating DEGs when comparing different magnitudes of shear. DEGs identified in a 1 dynes/cm 2 vs 0 dynes/cm 2 analysis are shown in green, DEGs identified in a 4 dynes/cm 2 vs 0 dynes/cm 2 are shown in orange, and identified in a 10 dynes/cm 2 vs 0 dynes/cm 2 are shown in purple. DEG, differentially expressed gene; EC, endothelial cell; HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell; NS, not significant; PCA, principal component analysis; RNA-seq, RNA sequencing.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques: RNA Sequencing, Expressing, Shear, Control

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: (A) A custom gene set of endothelial genes (see ) was used for DEG analysis to focus on endothelial-related genes and is displayed as a volcano plot of DEGs. Log 2 fold change (FC) is expressed as the ratio of expression of shear stress samples relative to control samples. The horizontal dashed line demarcates P = 10e-20, and the vertical dashed line represents a FC cutoff of 1.5 or −1.5. (B) Correlation plots and linear regression analysis examining the expression of known shear stress-dependent genes KLF2 , KLF4 , and NOS3 with shear stress demonstrate increased gene expression with increased shear stress across all EC types (HUVECs are shown in purple, HPAECs in green, and HMVECs in pink). Linear regression analysis was conducted and representative R 2 values and P values are shown. (E) A custom set of endothelial genes organized in gene sets named “atheroprotective,” “atheroprone,” “cytoskeletal,” or “shear” was used to generate a heatmap analysis. DEG, differentially expressed gene; EC, endothelial cell; EndoMT, endothelial-to-mesenchymal transition; HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell; NS, not significant.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques: Expressing, Shear, Control, Gene Expression

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: Heatmap of angiogenic genes. Heatmap analysis of all genes associated with the gene ontology biological process GO:0001525 (angiogenesis) as of 2024 is displayed. A list of genes can be found at https://amigo.geneontology.org/amigo/term/GO:0001525 . Overall, shear stress demonstrates a significant change in the expression of angiogenic genes. Similar to earlier heatmaps, there is some heterogeneity of up/downregulation of certain genes when comparing HMVECs vs HPAECs vs HUVECs. HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques: Shear, Expressing

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: Analysis of custom gene sets relating to hemostasis and von Willebrand factor (VWF). (A) A custom gene set of hemostatic endothelial genes was analyzed via heatmap analysis for transcriptional change associated with shear stress. Some genes, such as F2R and F3 , appear to have significant changes under conditions of shear stress. (B) Volcano plot of statistically significant differentially expressed genes in the hemostatic gene set. (C) Correlation plot of VWF expression with shear stress magnitude. VWF appears to be downregulated under conditions of flow in 5B, and the correlation plot generated in 5C demonstrates an overall pattern of decreasing VWF expression with increasing shear stress. Linear regression analysis was conducted and representative R 2 values and P values are shown. (D) Heatmap analysis of genes associated with VWF changes in GWASs. Based on the finding of decreased VWF expression with increasing shear stress, we evaluated a set of genes identified in VWF-related GWASs. EC, endothelial cell; FC, fold change; GWAS, genome-wide association study; HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques: Shear, Expressing, Generated, GWAS

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Vascular-type heterogeneity is associated with differential gene expression profiles of endothelial cells under shear stress

doi: 10.1016/j.rpth.2025.102894

Figure Lengend Snippet: Correlation coefficients of genome-wide association study-identified genes associated with von Willebrand factor levels. Spearman coefficients are displayed for respective vascular-type endothelial cells in the heatmap. The legend to the right displays the strength of the Spearman coefficient ranging from purple (low) to yellow (high). HMVEC, human microvascular endothelial cell; HPAEC, human pulmonary arterial endothelial cell; HUVEC, human umbilical vein endothelial cell.

Article Snippet: ECs were sustained in EGM-2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKit medium (Lonza) supplemented with 10% fetal bovine serum.

Techniques: GWAS