Journal: Nature Communications

Article Title: Post-myocardial infarction heart failure dysregulates the bone vascular niche

doi: 10.1038/s41467-021-24045-4

Figure Lengend Snippet: a scRNA-seq of mice bone marrow vascular niche 7 (d7) and 28 days (d28) after myocardial infarction as well as of controls (d0). Clustered cells from the three-time points are displayed in t-SNE plots. Emcn expressing cells are colored. b -left Representation of the percentage of Emcn expressing cells of the lineage depleted, CD31 expressing population. b -right Emcn and ( c ) Il1b and Myc average expression. d Representation of the ten most significant upregulated terms at d7 revealed by gene ontology analysis when comparing d0 to d7. e Immunostaining of longitudinal femur sections at d1. IL-1β immuno-signal is enriched in type H vessels one day after the ischemic insult. f -up RT-qPCR analysis of MYC expression in HUVEC cells after 1 h IL-1β treatment. N = 4. Data are shown as mean ± SEM. P -value was calculated by a two-tailed Mann–Whitney test. f -down Analysis of MYC activation in HUVEC cells after 1 h IL-1β treatment. N = 3 independent experiments with two technical replicates. Data are shown as fold-change relative to control. g Analysis of Caspase1 activation in HUVECs after 1 h IL-1β treatment or nigericin as a positive control. Left, Gating strategy. Right, quantification. N = 4. Data are shown as mean ± SEM. P -value was calculated by a two-tailed Mann–Whitney test. h Analysis of human MYC expression in isolated liver endothelial cells by RT-qPCR. N = 2 for both groups. Data are shown as mean ± SEM. i Immunostaining of longitudinal sections through the femur. The length of type H vessels (indicated by the dashed line, endomucin in red, CD31 in green, and DAPI in blue) is reduced in Myc EC-OE mice when compared to controls. Tamoxifen was given at 8 weeks and analysis performed 4 weeks later, N = 6 for both groups. Data are shown as mean ± SEM. P -value was calculated by unpaired, two-tailed Student’s t -test.

Article Snippet: HUVEC cells were seeded in 6 well cell culture plates (657160; Greiner Bio-One) and stimulated after 24 h with Nigericin (10 µM; N7143; Sigma Aldrich) or IL-1β (100 ng/mL; 201-LB; R&D Systems) for 3 h at 37 °C.

Techniques: Expressing, Immunostaining, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Activation Assay, Positive Control, Isolation

Journal: Cancer research

Article Title: A tissue-engineered 3D microvessel model reveals the dynamics of mosaic vessel formation in breast cancer

doi: 10.1158/0008-5472.CAN-19-1564

Figure Lengend Snippet: Frames from a representative (A) phase contrast and (B) fluorescence time-lapse movie of a ROSAmTmG; MMTV-PyMT tumor organoid and a HUVEC microvessel (GFP). Insets of phase (A’) and fluorescent (B’) time-lapse images showing a tumor cluster shedding into the vessel lumen under continuous flow (yellow arrows). The flow rate is kept at 1mL/hour and shear stress at 4 dynes/cm2. Direction of flow is from left to right. Scale bars: 100 μm.

Article Snippet: 2D co-culture and staining HUVEC-GFP cells were seeded in 24 well glass bottom plates (662892, Greiner Bio-one) and incubated for 24h in a humidified environment with 5% CO2 at 37°C.

Techniques: Fluorescence

Journal: Cancer research

Article Title: A tissue-engineered 3D microvessel model reveals the dynamics of mosaic vessel formation in breast cancer

doi: 10.1158/0008-5472.CAN-19-1564

Figure Lengend Snippet: (A) Schematic representation of workflow for isolating and imaging organoids in parallel tumor-vessel device. (B) Top- and end-view of microvessel device fabrication. To fix the tumor-vessel distance, a second cylindrical template rod is added into the device, with the collagen solution surrounding both rods. Tumor organoids at a high concentration are injected into one inlet and then the rod is slowly removed, allowing the introduction of the tumor/collagen gel suspension into the cylindrical channel. After the tumor/collagen solution gels, the second rod is removed, resulting in a bare channel which is perfused with media and coated with fibronectin. Endothelial cells are then seeded, resulting in a confluent 3D microvessel. The perfusion system is by gravity flow by differential pressure. Flow is 1 mL/h and shear stress at 4 dynes/cm2. (C) Schematic of the mid-plane of the tumor and vessel. (D) Fluorescence image of a ROSAmTmG; MMTV-PyMT tumor organoid-filled channel and a HUVEC microvessel (GFP), focusing at mid-plane. (E) Frames from a fluorescent time-lapse movie of a collective tumor strand that integrates into the vessel wall, forming a mosaic vessel. The tumor cells are exposed to flow, which is kept at 1mL/hour and shear stress at 4 dynes/cm2. Scale bars: (D) 200 μm, (E) 100 μm.

Article Snippet: 2D co-culture and staining HUVEC-GFP cells were seeded in 24 well glass bottom plates (662892, Greiner Bio-one) and incubated for 24h in a humidified environment with 5% CO2 at 37°C.

Techniques: Imaging, Concentration Assay, Injection, Fluorescence

Journal: Cancer research

Article Title: A tissue-engineered 3D microvessel model reveals the dynamics of mosaic vessel formation in breast cancer

doi: 10.1158/0008-5472.CAN-19-1564

Figure Lengend Snippet: Frames from a representative (A) phase contrast and (B) fluorescence time-lapse movie of a ROSAmTmG; MMTV-PyMT tumor organoid and a HUVEC microvessel (GFP). Insets of phase (A’) and fluorescent (B’) time-lapse images showing a tumor cluster shedding into the vessel lumen under continuous flow (yellow arrows). The flow rate is kept at 1mL/hour and shear stress at 4 dynes/cm2. Direction of flow is from left to right. Scale bars: 100 μm.

Article Snippet: HUVEC-GFP cells were seeded in 24 well glass bottom plates (662892, Greiner Bio-one) and incubated for 24h in a humidified environment with 5% CO2 at 37°C.

Techniques: Fluorescence

Journal: Cancer research

Article Title: A tissue-engineered 3D microvessel model reveals the dynamics of mosaic vessel formation in breast cancer

doi: 10.1158/0008-5472.CAN-19-1564

Figure Lengend Snippet: (A) Schematic representation of workflow for isolating and imaging organoids in parallel tumor-vessel device. (B) Top- and end-view of microvessel device fabrication. To fix the tumor-vessel distance, a second cylindrical template rod is added into the device, with the collagen solution surrounding both rods. Tumor organoids at a high concentration are injected into one inlet and then the rod is slowly removed, allowing the introduction of the tumor/collagen gel suspension into the cylindrical channel. After the tumor/collagen solution gels, the second rod is removed, resulting in a bare channel which is perfused with media and coated with fibronectin. Endothelial cells are then seeded, resulting in a confluent 3D microvessel. The perfusion system is by gravity flow by differential pressure. Flow is 1 mL/h and shear stress at 4 dynes/cm2. (C) Schematic of the mid-plane of the tumor and vessel. (D) Fluorescence image of a ROSAmTmG; MMTV-PyMT tumor organoid-filled channel and a HUVEC microvessel (GFP), focusing at mid-plane. (E) Frames from a fluorescent time-lapse movie of a collective tumor strand that integrates into the vessel wall, forming a mosaic vessel. The tumor cells are exposed to flow, which is kept at 1mL/hour and shear stress at 4 dynes/cm2. Scale bars: (D) 200 μm, (E) 100 μm.

Article Snippet: HUVEC-GFP cells were seeded in 24 well glass bottom plates (662892, Greiner Bio-one) and incubated for 24h in a humidified environment with 5% CO2 at 37°C.

Techniques: Imaging, Concentration Assay, Injection, Fluorescence