Journal: Bioactive Materials

Article Title: A catalytically active and recyclable bioelastomer inspired by metalloenzymes

doi: 10.1016/j.bioactmat.2026.02.053

Figure Lengend Snippet: Cytocompatibility of Cu-PIAS. ( A ) Viability, as measured by ATP content, of human umbilical vein endothelial cells cultured with media containing extracts of 15-Cu-PIAS, Latex (Positive), or PCL (Negative) over three days. ( B ) Representative images of cell morphology after 24 h exposure to material extract-containing media, visualized by Calcein AM (Green) staining. Additional staining with ethidium homodimer-1 (red) indicates the presence of non-viable cells.

Article Snippet: Briefly, endothelial growth medium supplemented with 5% fetal bovine serum, growth factors, ascorbic acid, and hydrocortisone (Promocell C-22121) was incubated at 37 °C for 72 h under agitation alone, or in the presence of 15-Cu-PIAS (6 cm 2 /mL), PCL (6 cm 2 /mL), or latex (3 cm 2 /mL) sheets cut into several 6 mm × 6 mm squares.

Techniques: Cell Culture, Staining

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericyte secretions mediate the inhibition of TNBC cell adhesion to the BBB. (A) Experimental workflow for adhesion assay. All BBB experiments were performed using ECGMV2 medium with supplements for all conditions from day −6 to the end of the experiment. The astrocyte medium was renewed 24 h before the adhesion experiment with the same ECGMV2 medium. Endothelial cells (ECs) and brain pericytes (hBPs) were co‐cultured for 5 days, and inserts containing brain‐like endothelial cells (hBLECs) were then either maintained with hBPs or transferred to wells without hBPs. After 24 hours, and prior to MDA‐MB‐231 cells seeding, inserts were subjected to the following conditions: maintained without hBPs (hBLEC 24h); transferred to hBPs previously cultured alone for 24 hours (hBLEC 24h then hBP); transferred to wells without hBPs but containing either culture medium (hBLEC) or CM from co‐culture (hBLEC + CM coc ); transferred to wells with astrocytes (hBLEC + A) or maintained in co‐culture with hBPs (hBLEC + hBP). (B–G), Quantification (B, D, F) of MDA‐MB‐231 cell adhesion to hBLECs after 3 h of incubation and representative images (C, E, G) of adhered MDA‐MB‐231 cells (green) under the different conditions. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using one‐way ANOVA followed by Tukey's test (B), Welch's ANOVA followed by Dunnett's T3 test (D) and Kruskal–Wallis test followed by Dunn's test (F). CM coc = conditioned medium from 24 h hBLECs and hBPs coculture, A = astrocytes, hBP mono = 24 h hBPs monoculture. Scale bar = 750 µm. BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; ECs, endothelial cells; hBLECs, human brain‐like endothelial cells; ns, non‐significant; hBPs, human brain pericytes; TNBC, triple‐negative breast cancer. **** p ≤ 0.0001; * p ≤ 0.05.

Article Snippet: Differentiated ECs were cultured in 100‐mm Petri dishes (Corning) coated with gelatin (Sigma‐Aldrich), in EC growth medium MV 2 (ECGMV2 + Supplement mix C‐39226, PromoCell) and gentamicin (50 μg/ml; Biowest). hBPs (female) were provided by Dr. Fumitaka Shimizu and Pr.

Techniques: Inhibition, Cell Adhesion Assay, Cell Culture, Co-Culture Assay, Incubation

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes limit TNBC cell adhesion to the BBB under hypoxic conditions. (A) Experimental workflow for adhesion assay in hypoxic conditions. All BBB experiments were performed using ECGMV2 medium with supplements for all conditions from day −6 to the end of the experiment. After 5 days of co‐culture with brain pericytes (hBPs), inserts containing brain‐like endothelial cells (hBLECs) were exposed to hypoxia for 24 h in the presence or absence of hBPs. Parallel conditions maintained in normoxia served as controls. (B–E) Quantification (B, D) of MDA‐MB‐231 cell adhesion to hBLECs after 3 h of incubation and representative images (C, E) of adhered MDA‐MB‐231 cells (green) under the different conditions. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using Welch's ANOVA followed by Dunnett's T3 test (B) and Kruskal‐Wallis test followed by Dunn's test (D). hBP‐CM mono = conditioned medium from 24 h hBPs monoculture. Scale bar = 750 μm. BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; hBLECs, human brain‐like endothelial cells; hBPs, human brain pericytes; ns, non‐significant; TNBC, triple‐negative breast cancer. *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05.

Article Snippet: Differentiated ECs were cultured in 100‐mm Petri dishes (Corning) coated with gelatin (Sigma‐Aldrich), in EC growth medium MV 2 (ECGMV2 + Supplement mix C‐39226, PromoCell) and gentamicin (50 μg/ml; Biowest). hBPs (female) were provided by Dr. Fumitaka Shimizu and Pr.

Techniques: Cell Adhesion Assay, Co-Culture Assay, Incubation

Journal: Journal of Cell Communication and Signaling

Article Title: Human brain pericytes protect the blood–brain barrier from triple‐negative breast cancer cells while promoting tumor aggressiveness

doi: 10.1002/ccs3.70070

Figure Lengend Snippet: Brain pericytes maintain BBB integrity in the presence of TNBC cells under hypoxic conditions and supplement deprivation. (A) After 5 days of co‐culture with brain pericytes (hBPs) in ECGMV2 medium with supplements, inserts containing brain‐like endothelial cells (hBLECs) co‐cultured with hBPs were exposed to hypoxia for 16 h following medium replacement with ECGMV2 basal medium (without supplements). Then, endothelial permeability (Pe) to Lucifer Yellow was assessed after 3 h of incubation with MDA‐MB‐231 cells, in the presence or absence of hBPs, under normoxic and hypoxic conditions. (B) Permeability values (expressed in x10 −3 cm/min ± SD) are summarized in a table. (C) Representative images of endothelial Claudin‐5 (magenta) immunostaining in the absence or presence of hBPs after 3 h of incubation with MDA‐MB‐231 cells (green) under hypoxic conditions in basal medium. Data are obtained from three independent experiments, with three technical replicates per condition. Statistical analyses were performed using Welch's ANOVA followed by Dunnett's T3 test. MDA = MDA‐MB‐231 cells. Scale bars = 300 μm (10X) and 100 μm (40X). BBB, blood–brain barrier; ECGMV2, EC Growth Medium MV 2; hBLECs, human brain‐like endothelial cells; hBPs, human brain pericytes; ns, non‐significant; TNBC, triple‐negative breast cancer. *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05.

Article Snippet: Differentiated ECs were cultured in 100‐mm Petri dishes (Corning) coated with gelatin (Sigma‐Aldrich), in EC growth medium MV 2 (ECGMV2 + Supplement mix C‐39226, PromoCell) and gentamicin (50 μg/ml; Biowest). hBPs (female) were provided by Dr. Fumitaka Shimizu and Pr.

Techniques: Co-Culture Assay, Cell Culture, Permeability, Incubation, Immunostaining

Journal: JACC: Basic to Translational Science

Article Title: Endothelial Senescence Drives Deleterious Endothelial-Adipocyte Cross-Talk in Patients With Heart Failure and Type 2 Diabetes

doi: 10.1016/j.jacbts.2026.101527

Figure Lengend Snippet: SATMVECs From Patients With HFDM Exhibit a Senescent Phenotype (A) Subcutaneous adipose tissue microvascular endothelial cell (SATMVEC) size (n = 8, 7). (B) SATMVEC senescence histochemical senescence-associated β-galactosidase (b-gal) staining with illustrative phase and fluorescent imaging (n = 16, 12). (C) SATMVEC proliferation using the Click-iT EdU imaging kit (n = 19, 14). (D) SATMVEC mitochondrial adenosine triphosphate production (n = 12, 11). (E) Messenger RNA expression of RB1 in SATMVEC plotted with jittered data points. Significance is Bonferroni adjusted (n = 56, 30). (F) SATMVEC tube forming in Matrigel with illustrative phase images (n = 15, 9). (G) Secreted interleukin (IL)–6 in SATMVEC conditioned medium standardized to total protein concentration quantified using Luminex (Sigma-Aldrich) (n = 16, 17). (H) Secreted IL-8 in SATMVEC conditioned medium standardized to total protein concentration quantified using Luminex (n = 16, 16). Data are presented as mean ± SEM, with individual data points representing biological replicates, unless otherwise indicated. Violin plots show the distribution of individual values. P values for comparisons between the heart failure (HF) and HF with type 2 diabetes mellitus (HFDM) groups were calculated using Student’s t -test or the Mann-Whitney U test as appropriate according to data distribution. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. AU = arbitrary units; HPF = high power field; OCR = oxygen consumption rate; TPM = transcripts per million.

Article Snippet: Once fully differentiated, adipocytes were cocultured for 24 hours with SATMVECs (passage 4) grown to confluence on 0.4-μm Transwell inserts (Corning) in microvascular endothelial cell growth medium (PromoCell).

Techniques: Staining, Imaging, RNA Expression, Protein Concentration, Luminex, MANN-WHITNEY

Journal: JACC: Basic to Translational Science

Article Title: Endothelial Senescence Drives Deleterious Endothelial-Adipocyte Cross-Talk in Patients With Heart Failure and Type 2 Diabetes

doi: 10.1016/j.jacbts.2026.101527

Figure Lengend Snippet: Cross-Talk Between HFDM SATMVECs and Adipocytes Induces a Proinflammatory, Metabolically Dysregulated Phenotype (A) Illustrative diagram showing set up of coculture experiments between SATMVECs and adipocytes. (B) Expression of adipocyte IL-6 messenger RNA (mRNA) following 24-hour coculture with SATMVECs (n = 17, 16). (C) Expression of adipocyte ADIPOQ mRNA following 24-hour coculture with SATMVECs (n = 17, 15). (D) Expression of adipocyte GLUT4 mRNA following 24-hour coculture with SATMVECs (n = 15, 15). (E) Uptake of 2-NBD-glucose in adipocytes cocultured with SATMVECs for 24 hours with illustrative images (n = 16, 16). (F) Quantification and representative immunoblot of total and cell-surface GLUT4 protein in adipocytes cocultured with SATMVECs from HF and HFDM patients for 24 hours (n = 9, 10). (G) Correlation between relative membrane-bound GLUT4 expression in adipocytes and patient HbA 1c levels. (H) Correlation between relative membrane-bound GLUT4 expression in adipocytes and patient age. Data are presented as mean ± SEM, with individual data points representing biological replicates. P values for comparisons between the HF and HFDM groups were calculated using Student’s t -test or the Mann-Whitney U test as appropriate on the basis of data distribution. P values for correlation analyses in G and H were calculated using Pearson’s correlation coefficient. ∗ P < 0.05. GAPDH = glyceraldehyde-3-phosphate dehydrogenase; MV = microvascular; PDM = preadipocyte differentiation media; WCL = whole-cell lysate; other abbreviations as in and .

Article Snippet: Once fully differentiated, adipocytes were cocultured for 24 hours with SATMVECs (passage 4) grown to confluence on 0.4-μm Transwell inserts (Corning) in microvascular endothelial cell growth medium (PromoCell).

Techniques: Metabolic Labelling, Expressing, Western Blot, Membrane, MANN-WHITNEY