microvascular endothelial cell growth medium  (PromoCell)

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

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