Journal: Materials Today Bio

Article Title: Redirecting the route: Monocyte-mediated delivery of oHSV-1 across a human BBB-on-chip model

doi: 10.1016/j.mtbio.2025.102458

Figure Lengend Snippet: Microfluidic BBB-on-chip design and validation. A. Schematic representation of the BBB-on-chip, seeded with endothelial cells on the blood side and pericytes and astrocytes on the brain side, equipped with three microwells for 3D GBM spheroid culture (GBM spheroids area). B. Image of the assembled BBB-on-chip showing the upper channel (yellow) mimicking the vascular compartment and main perfusion conduit, and the lower channels (red) leading to the tumor compartment (scale bar = 2 cm). C. Transendothelial electrical resistance (TEER) measurements recorded daily across the membrane between the upper and lower channels and supporting the BBB. D. Permeability coefficients calculated from the diffusion of 40, 70, and 150 kDa FITC-dextrans through the assembled BBB-on-chip in the following conditions: i. No cells (control, membrane without cells), ii. Blood side (membrane + HUVECs endothelial monolayer), and iii. BBB complete (membrane with the three-culture of HUVECs on the blood side and pericytes and astrocytes on the brain side). E . IF staining of the BBB-on-chip model on day 7. On the blood side compartment , endothelial tight junction protein ZO-1 (green) and the endothelial marker CD31 (green) are expressed, confirming the formation of a continuous endothelial layer. On the brain side , astrocytes are identified by GFAP expression (magenta), indicating appropriate localization. DAPI (blue) highlights cell nuclei in both compartments. Scale bar = 50 μm. Values are expressed as median±SEM from at least 3 independent experiments. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 vs control. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The assembled BBB-on-chip devices were bound to glass coverslips via further plasma treatment and sterilized with 70 % ethanol followed by UV exposure for 60 min. BBB-on-chip cells seeding and culturing: Human umbilical vein endothelial cells (HUVECs), brain vascular pericytes (hBVPs), and immortalized astrocytes (hAs) were obtained from Innoprot (Derio, Spain).

Techniques: Biomarker Discovery, Membrane, Permeability, Diffusion-based Assay, Control, Staining, Marker, Expressing

Journal: The European Journal of Neuroscience

Article Title: VEGF‐E Attenuates Injury After Ischemic Stroke by Promoting Reparative Revascularization

doi: 10.1111/ejn.70114

Figure Lengend Snippet: VEGF‐E‐stimulated brain endothelial cells promote perivascular cell mobility under ischemia and reperfusion‐like condition. (a) Schematic illustration of the experimental design of iHBEC exposed to OGD and reperfusion followed by stimulation with vehicle (VEH) or VEGF‐E (100 ng/mL) for 24 h. (b) Representative Western blot images of phosphorylated (p) and total (t) ERK1/2 expression in iHBEC stimulated with vehicle (VEH) or VEGF‐E. Analysis of (c) p‐ERK1 and (d) t‐ERK1 expression as well as (e) p‐ERK1/t‐ERK1 ratio (i.e., pathway activation). Analysis of (f) p‐ERK2 and (g) t‐ERK2 expression as well as (h) p‐ERK2/t‐ERK2 ratio (i.e., pathway activation) in iHBEC stimulated with VEH and VEGF‐E after OGD. (i) Representative Western blot images of phosphorylated (p) and total (t) P38 expression. Analysis of (j) p‐P38 and (k) t‐P38 expression as well as (l) p‐P38/t‐P38 ratio (i.e., pathway activation) in iHBEC stimulated with VEH and VEGF‐E after OGD. (m) Schematic illustration of the experimental design of wound healing assay performed on HBVP after stimulation with VEH, VEGF‐E, or the conditioned medium (CM) of VEGF‐E‐treated iHBEC. (n) Representative brightfield images of the wound in VEH, VEGF‐E, or CM‐stimulated HBVP at baseline, 24 h, or 48 h. Analysis of wound closure of VEH, VEGF‐E, or CM‐treated HBVP at (o) 24 h and (p) 48 h, represented as percent of control (baseline, 0 h). Data are boxplot with min/max (n = 3–4 independent experiments/condition). * p < 0.05/** p < 0.01/*** p < 0.001 compared to control (c‐e, j, l, unpaired two‐tailed t ‐test; o, p, one‐way ANOVA). Abbreviations: H, hours; iHBEC, immortalized human brain endothelial cells; OGD, oxygen and glucose‐deprived; VEGF, vascular endothelial growth factor; VEH, vehicle.

Article Snippet: Moreover, primary human brain vascular pericytes (HBVP) (ScienCell; 1200; 33634) were used to investigate the action of VEGF‐E on factor release from iHBEC cells.

Techniques: Western Blot, Expressing, Activation Assay, Wound Healing Assay, Control, Two Tailed Test

Journal: Journal of Tissue Engineering

Article Title: Construction of functional tissue-engineered microvasculatures using circulating fibrocytes as mural cells

doi: 10.1177/20417314251315523

Figure Lengend Snippet: Both CFs and HBVPs inhibited the proliferation of VECs in 2D culture dishes and inhibited tube extension in a collagen gel. (a) VECs were labelled with the Vybrant ® DiI cell tracer. HBVPs and CFs were either cultured alone or cocultured with VECs in culture dishes for 24 h and were then observed via fluorescence microscopy. (b) VECs were either cultured alone or cocultured with CFs (VECs + Fs) or HBVPs (VECs + HBVPs) in transwell plates and processed for proliferation assays with a CellTiter-Glo ® Luminescent Cell Viability Assay Kit (Cat # G7570, Promega). *Indicates p < 0.05 versus VECs + CFs, and # indicates p < 0.05 versus VECs + HBVPs. (c), CFs or HBVPs were cocultured with VECs in transwell plates (VECs + CFs and VECs + HBVPs) and then processed as described in (b). *Indicates p < 0.05 versus VECs + CFs. (d and e) VECs were either cultured alone in a collagen gel or cocultured with CFs or HBVPs for 3 (d) or 10 days (e). The VECs shown in the upper panel were infected with ad-GFP, and the CFs or HBVPs were labelled with the Vybrant ® DiI cell tracer; the collagen gel was processed for toluidine blue staining (middle panel) or SEM (lower panel). (f) The lumen areas in (d) and (e) were quantified. *Indicates p < 0.05 versus VECs, and # indicates p < 0.05 versus VECs + CFs.

Article Snippet: Human umbilical vascular endothelial cells (HUVECs; Cat. No. 1000, ScienCell Research Laboratories, USA), human bone marrow-derived mesenchymal stem cells (BMSCs; Cat. No. 7500, ScienCell Research Laboratories, USA) and human brain vascular pericytes (HBVPs; Cat. No. 1200, ScienCell Research Laboratories, USA) were cultured according to the supplier’s protocol.

Techniques: Cell Culture, Fluorescence, Microscopy, Cell Viability Assay, Infection, Staining

Journal: Journal of Tissue Engineering

Article Title: Construction of functional tissue-engineered microvasculatures using circulating fibrocytes as mural cells

doi: 10.1177/20417314251315523

Figure Lengend Snippet: GO functional analysis of differentially expressed proteins. (a) Proteins were isolated from CFs and HBVPs and processed for proteomics analysis, and GO functional analysis was performed with the proteins upregulated in CFs via the Function Annotation tool in Metascape v3.5.20240101. (b) The same experimental procedure described in (a) was performed, and analysis was conducted with the downregulated proteins. (c) The same experimental procedure described in (a) was performed, but the proteins were isolated from CFs cocultured with VECs (CFs-VECs) and from CFs cultured alone (CFs), and the proteins downregulated in CFs-VECs were analysed. (d) The same experimental procedure described in (c) was performed, and the proteins upregulated in CFs-VECs were analysed. (e) Proteins were isolated from CFs cocultured with VECs (CFs-VECs) and HBVPs cultured with VECs (pericyte-VECs), and the proteins upregulated in CFs-VECs were analysed. (f) The same experimental procedure described in (e) was performed, and the proteins downregulated in CFs-VECs were analysed.

Article Snippet: Human umbilical vascular endothelial cells (HUVECs; Cat. No. 1000, ScienCell Research Laboratories, USA), human bone marrow-derived mesenchymal stem cells (BMSCs; Cat. No. 7500, ScienCell Research Laboratories, USA) and human brain vascular pericytes (HBVPs; Cat. No. 1200, ScienCell Research Laboratories, USA) were cultured according to the supplier’s protocol.

Techniques: Functional Assay, Isolation, Cell Culture

Journal: Journal of Tissue Engineering

Article Title: Construction of functional tissue-engineered microvasculatures using circulating fibrocytes as mural cells

doi: 10.1177/20417314251315523

Figure Lengend Snippet: Tissue-engineered capillary-like structures connected to the host circulation. (a) Schematic diagram showing the experimental design for (b) and (c). The colours in the chart do not represent the colours of fluorescence staining in the experiment. VECs and CFs were cultured alone or cocultured in a collagen gel for 7 days in vitro and were then injected intramuscularly into nude mice. Samples were harvested 7 days later for follow-up testing. CFs and HBVPs were labelled with the Vybrant ® DiI cell tracer. (b) Qtracker ® 655 quantum dots (Cat #. Q21021MP, Thermo Fisher Scientific, US) were injected into the tail vein immediately before sample harvesting; the dots were retained as labels in the blood vessels. The samples were observed via confocal microscopy. (c) Samples were harvested and processed for TEM. M indicates Matrigel, L indicates the lumen, and the black arrowhead indicates the basement membrane. (d) Experimental design for the calvarial defect model in nude rats. The schematic shows the experimental design for (e)-(g). (e) Micro-CT scans acquired 8 weeks after calvarial defect repair. (f) Images of Masson’s trichrome-stained samples. (g) Immunofluorescence images of the samples. Both the CFs and HBVPs expressed CD34 and CD13 (orange cells in the second and third rows). The BMSCs expressed only CD13 (red cells), the VECs expressed only CD34 (green cells), and the nuclei were labelled with DAPI (blue).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs; Cat. No. 1000, ScienCell Research Laboratories, USA), human bone marrow-derived mesenchymal stem cells (BMSCs; Cat. No. 7500, ScienCell Research Laboratories, USA) and human brain vascular pericytes (HBVPs; Cat. No. 1200, ScienCell Research Laboratories, USA) were cultured according to the supplier’s protocol.

Techniques: Fluorescence, Staining, Cell Culture, In Vitro, Injection, Confocal Microscopy, Membrane, Micro-CT, Immunofluorescence