hcmec d3 cells  (Inserm Transfert)

Journal: Advanced Science

Article Title: 3D Bioprinted Blood–Brain–Tumor Barrier Spheroid‐Based In Situ Evaluation of Anticancer Compounds Against Glioblastoma: ROS‐Producing Ruthenium as a Potential Therapeutic Candidate

doi: 10.1002/advs.78150

Figure Lengend Snippet: Cytotoxicity profiling and Transwell permeability assessment of test compounds across hCMEC/D3 and U‐87 MG cell models. (A1–A4) Cell viability of hCMEC/D3 human cerebral microvascular endothelial cells following 24 h treatment with Rh123 (A1), Ru (A2), Osi (A3), or TMZ (A4), assessed by CellTiter‐Glo luminescent cell viability assay. (B1–B3) Cell viability of U‐87 MG glioblastoma cells following 48 h treatment with Ru (B1), Osi (B2), or TMZ (B3), assessed by MTT assay. (C1–C4) Cumulative apical‐to‐basolateral transport profiles of Rh123 (C1), Ru (C2), Osi (C3), and TMZ (C4) across hCMEC/D3 monolayers in the presence (Ver+) or absence (Ver−) of the P‐glycoprotein inhibitor verapamil (Ver, 100 µM), measured at 15‐min intervals over 60 min. (D1–D4) Apparent permeability coefficients (P app ) of Rh123 (D1), Ru (D2), Osi (D3), and TMZ (D4) in the presence (Ver+) or absence (Ver−) of verapamil. ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant. Rh123, Rhodamine‐123; Ru, ruthenium compound; Osi, osimertinib; TMZ, temozolomide; Ver, verapamil. Data in (A1–A4) and (B1–B3) are presented as mean ± SD of n = 3 replicate wells per concentration seeded in parallel; data in (C1–C4) and (D1–D4) are presented as mean ± SD of n = 3 Transwell inserts seeded in parallel per condition. Statistical significance was assessed by ordinary one‐way ANOVA followed by Tukey's multiple‐comparisons test, using a two‐sided alpha of 0.05. Analyses were performed using GraphPad Prism 11.0.2.

Article Snippet: Human cerebral microvascular endothelial cells (hCMECs/D3; Cedarlane, Canada; RRID: CVCL_0M40) were cultured in Endothelial Growth Medium 2 (EGM2; Lonza Bioscience, Switzerland), containing human epidermal growth factor, hydrocortisone, GA 1000, FBS, vascular endothelial growth factor (VEGF), human fibroblastic growth factor (hFGF‐B), R 3 IGF‐I, ascorbic acid, and heparin (Lonza).

Techniques: Profiling, Permeability, Cell Viability Assay, MTT Assay, Ruthenium, Concentration Assay

Journal: Virulence

Article Title: Integrated transcriptomic and proteomic analysis reveals inflammatory activation and blood-brain barrier disruption during meningitis-associated extraintestinal pathogenic Escherichia coli infection

doi: 10.1080/21505594.2026.2670939

Figure Lengend Snippet: RS218 infection induces cell death and tight junction disruption in vitro . (A) Relative LDH release in RS218-infected hCMEC/D3 cells pretreated with calcium chelator BAPTA-AM (10 μM), necroptosis inhibitor Necrostatin-1 (10 μM), caspase-1 inhibitor VX-765 (10 μM), or caspase-3 inhibitor Z-DEVD-FMK (20 μM). (B) LDH release in BV2 cells and murine peritoneal macrophages (PMs) infected with RS218 at MOI = 0.1, 1, 10, and 100 for 1 h and 2 h. (C) Immunofluorescence staining of tight junction protein ZO-1 in hCMEC/D3 cells following RS218 infection. Representative images show DAPI (blue), ZO-1 (green), and merged channels in control and infected cells. Scale bars, 50 μm. Bar graphs show ZO-1 positive area and fluorescence intensity in infected cells relative to controls ( n = 3).

Article Snippet: For inhibitor experiments, hCMEC/D3 cells were pretreated for 1 h with BAPTA-AM (10 μM), Necrostatin-1 (10 μM), VX-765 (10 μM), or Z-DEVD-FMK (20 μM) (all from MedChemExpress, USA) prior to RS218 infection.

Techniques: Infection, Disruption, In Vitro, Immunofluorescence, Staining, Control, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Establishment and characterization of an inflammatory tricellular BBB in vitro model. ( A ) Changes in TEER of hCMEC/D3 cells before and after LPS treatment, with LPS stimulation significantly reducing TEER levels. ( B ) Permeability assay using FITC-dextran, showing an approximately 2.6-fold increase in permeability following LPS stimulation, indicating barrier function impairment. ( C ) Immunofluorescence reveals MAP2⁺ neuronal processes extending along GFAP⁺ astrocytic end-feet, forming synapse-endfoot contacts. ( D ) Top-down view of cellular co-localization in the tricellular co-culture system, displaying neurons, astrocytes, and endothelial cells co-existing within the same field of view. ( E ) Schematic representation of the spatial distribution within the tricellular in vitro BBB model, illustrating the arrangement of different cells in the Transwell system. ( F ) VE-cadherin immunostaining shows changes in endothelial cell junction integrity before and after LPS treatment, with blurred junctional boundaries and reduced expression post-stimulation. Data are presented as mean ± SEM. Statistical significance in panels A and B was determined using a two-tailed unpaired Student’s t-test, **p < 0.01, ***p < 0.001

Article Snippet: The human cerebral microvascular endothelial cell line hCMEC/D3 (ATCC ® CRL-3245 TM ) was cultured at 37 °C in a humidified atmosphere containing 5% CO 2 using EndoGro TM -MV complete medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 1% L-glutamine.

Techniques: In Vitro, Permeability, Immunofluorescence, Co-Culture Assay, Immunostaining, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Enhanced targeted uptake of ICAM-1-NP@Irisin in inflammatory brain endothelial cells. ( A ) Confocal laser scanning micrographs: Representative fluorescence images of ICAM-1-NP@Irisin and NP@Irisin uptake in inflammatory hCMEC/D3 cells at 1 h, 3 h, and 6 h (red indicates NP fluorescence, blue indicates DAPI nuclear staining, scale bar 25 μm); the ICAM-1-NP@Irisin group exhibits stronger fluorescence signals with higher distribution density. ( B ) Quantitative analysis of fluorescence intensity: At all time points, intracellular fluorescence signals in the ICAM-1-NP@Irisin group are significantly higher than those in the NP@Irisin group (1 h: 1.4 ± 0.1 fold; 3 h: 1.8 ± 0.2 fold; 6 h: 2.1 ± 0.2 fold; data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001).

Article Snippet: The human cerebral microvascular endothelial cell line hCMEC/D3 (ATCC ® CRL-3245 TM ) was cultured at 37 °C in a humidified atmosphere containing 5% CO 2 using EndoGro TM -MV complete medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 1% L-glutamine.

Techniques: Fluorescence, Staining