Journal: International Journal of Molecular Sciences

Article Title: Ketone Bodies Promote Amyloid-β 1–40 Clearance in a Human in Vitro Blood–Brain Barrier Model

doi: 10.3390/ijms21030934

Figure Lengend Snippet: The effect of KBs on BLEC permeability. ( A ) Schematic representation of the human in vitro BBB model used for the KB study. This model is composed of an apical compartment in the filter containing endothelial cells and a basolateral compartment in the well with brain pericytes. After 5 days of co-culture, BLECs were incubated with various concentrations of KBs, AcAc, or βHB alone (5–20 mM) or in in combination (20 mM AcAc/20 mM βHB) for 48 h. Following KB treatment, the BLECs on the insert were transferred to a new plate for experiments. ( B ) The effects of KBs on cell viability were analyzed by MTT assay. ( C ) BLEC monolayer integrity was determined by measuring the endothelial lucifer yellow permeability (Pe LY ). Each bar represents the mean ± SEM relative to the control conditions (Pe LY = 0.92 ± 0.03 × 10 −3 cm∙min −1 ). The results are representative of three independent experiments performed in triplicate (*** p < 0.001). ( D ) Associated tight junction protein ZO-1 (green) and tight junction protein claudin-5 (red) staining were stained using immunofluorescence. Interruptions in the staining are indicated by white arrows. Nuclei were stained with Hoechst reagent and appear in blue. Scale bar: 50 µm.

Article Snippet: Endothelial cell medium (ECM; Sciencell, USA) supplemented with 5% heat-inactivated fetal calf serum (FCS, GIBCO, Life Technology SAS, Saint Aubin, France), 50 μg·mL −1 gentamicin (Biochrom GmbH, Germany), and 0.5% endothelial cell growth factor (Sciencell, USA) (ECM-5) was prepared and stored at 4 °C for a maximum of one week.

Techniques: Permeability, In Vitro, Co-Culture Assay, Incubation, MTT Assay, Control, Staining, Immunofluorescence

Journal: Acta Neuropathologica Communications

Article Title: Microembolus clearance through angiophagy is an auxiliary mechanism preserving tissue perfusion in the rat brain

doi: 10.1186/s40478-020-01071-9

Figure Lengend Snippet: Impedance measurements with ECIS after addition of microparticles to HUVECs. a Example tracing of an ECIS experiment after addition of different amounts of microspheres or fibrin clots to endothelial cells. Tracings are shown as normalized electrical resistance, where t 0 is the baseline electrical resistance of ~ 1 h prior to addition of microparticles. Baseline electrical resistance at 400 Hz was 1086 Ω (median; range 633–1513 Ω). The arrows indicate the time point used for quantification in b . b Quantification of electrical resistance at 400 Hz 2 h after addition of microparticles. N = 5 independent experiments done in triplo. Data are depicted as mean ± S.D. One-way ANOVA with Tukey–Kramer’s test for multiple comparisons

Article Snippet: Human umbilical vein endothelial cells (HUVECs; Lonza, Verviers, Belgium, passage 2–5) were grown to confluence on 5 μg/ml fibronectin-coated glass coverslips (MenzelTM, ThermoFisher, Amsterdam, The Netherlands) and kept in Endothelial Cell Medium (ECM) supplemented with singlequots (ScienCell Research Laboratories, Carlsbad, CA) at 5% CO 2 at 37 °C.

Techniques:

Journal: Clinical and Translational Medicine

Article Title: Targeting capacity, safety and efficacy of engineered extracellular vesicles delivered by transdermal microneedles to treat plasmacytoma in mice

doi: 10.1002/ctm2.70327

Figure Lengend Snippet: CD38‐EVs targeting tumour cells in vitro. PKH26‐labelled extracellular vesicles (EVs) and CD38‐EVs incubated with RPMI8226 cells (A) and U266 cells (B) for 4, 8 and 12 h, with cell fluorescence measured by flow cytometry (FCM). Kinetics of uptake of EVs and CD38‐EVs within 12 h by RPMI8226 cells (C) and U266 cells (D), measured by relative fluorescence units (RFU). (E) RPMI8226 and U266 cells were incubated with EVs and CD38‐EVs for 8 h and visualised using laser scanning confocal microscope (LSCM) (scale bars: 5 µm). PKH26‐labelled EVs and CD38‐EVs incubated with RPMI8226/HUVECs and RPMI8226/BMSCs (F) and U266/HUVECs and U266/BMSCs (G) co‐culture systems for 12 h, with cell fluorescence measured by FCM. Kinetics of uptake of EVs and CD38‐EVs by myeloma cells, human umbilical vein endothelial cells (HUVECs) and bone marrow stromal/stem cells (BMSCs) in RPMI8226/HUVECs and RPMI8226/BMSCs (H) and U266/HUVECs and U266/BMSCs (I) co‐culture systems, measured by RFU. Data are generated from three independent experiments.

Article Snippet: The HUVECs cell line was grown in ECM medium containing an endothelial cell growth medium bullet kit (Sciencell).

Techniques: In Vitro, Incubation, Fluorescence, Flow Cytometry, Microscopy, Co-Culture Assay, Generated