Journal: Advanced Science

Article Title: Pericytes Contribute to Dysfunction in a Human 3D Model of Placental Microvasculature through VEGF‐Ang‐Tie2 Signaling

doi: 10.1002/advs.201900878

Figure Lengend Snippet: Placental pericytes reduce microvessel growth and connectivity. a) Schematic diagram showing pericyte location in relation to microvessels in vivo. b) Confocal image of HPP‐cocultured with HUVEC fixed at day 5. Shown is a single XY plane and orthogonal projections demonstrating lumen (red) wrapped by HPPs (green), as indicated by white arrows. Nuclei were labeled with Dapi (blue). Scale bar is 200 µm. c) Schematic showing the various geometric measurements using binary projection images. d) Comparison of mean vessel area (EC coverage), branch length, and microvessel connectivity between HLF and HPP cocultures. Significant differences between parameters appear early on. e) Parameters are compared for HPP cocultures with (green) and without (gray) added VEGF+FGF. Shown is mean ± s.e.m. * P > 0.05 with t ‐test.

Article Snippet: GFP‐labeled HPP (microvascular) were acquired from Angioproteomie and were cultured in pericyte growth medium according to manufacturer's protocols.

Techniques: In Vivo, Labeling, Comparison

Journal: Advanced Science

Article Title: Pericytes Contribute to Dysfunction in a Human 3D Model of Placental Microvasculature through VEGF‐Ang‐Tie2 Signaling

doi: 10.1002/advs.201900878

Figure Lengend Snippet: A triculture model for increased microvessel connectivity. a) A triculture microvascular system perfused with fluorescently labeled beads. HUVEC—red, pericytes—green, 10 µm beads—magenta. b) Binary images from maximum intensity projections for co‐ and tricultures, as shown at day 5. c) Schedule for media change from full growth endothelial growth medium (EGM) to reduced serum basal medium (EBM). d) Representative flow cytometry density plots for HPP, HLF, and tricultures. e) Mean population of ECs and stromal cells for the co‐ and tricultures at day 5, as measured by flow cytometry. Three separate devices for each culture condition were used for measurement and repeated in n = 3 separate experiments. f) Microvessel parameters are compared between co‐ and tricultures. Shown is mean ± s.e.m. Significance is indicated by * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one‐way ANOVA and Tukey test.

Article Snippet: GFP‐labeled HPP (microvascular) were acquired from Angioproteomie and were cultured in pericyte growth medium according to manufacturer's protocols.

Techniques: Labeling, Flow Cytometry

Journal: Advanced Science

Article Title: Pericytes Contribute to Dysfunction in a Human 3D Model of Placental Microvasculature through VEGF‐Ang‐Tie2 Signaling

doi: 10.1002/advs.201900878

Figure Lengend Snippet: Pericytes influence PE‐affiliated cytokine expression and endothelial barrier function. a) Cytokine expression is shown for HPP, HLF, and triculture microvessel supernatants collected at day 5. HPPs result in increased PE‐associated cytokine expression, as indicated by the last row demonstrating those that are up (+) and down (−) regulated in PE. ND—no‐data in found. Here red values are high, blue low, and white are mid‐level (0.5). All cytokines were normalized to numbers between 1 and 0 based on maximum and minimum intensities from the cytokine array (Figure S4b, Supporting Information). b) Ang1/2 expression analyzed by ELISA for co‐ and tricultures, measured from pooled samples ( n = 5). c) Permeability of microvessels perfused with 10 kDa dextran (blue) at day 7 for co‐ and tricultures. Shown is mean ± s.e.m. Significance is indicated by * P < 0.05, ** P < 0.01, using t ‐test. d) Confocal images demonstrating perfusability of HLF cocultures and Tricultures, and lack of perfusability in HPP cocultures. HUVEC—red, 10 kDa dextran–blue. Scale bar is 200 µm.

Article Snippet: GFP‐labeled HPP (microvascular) were acquired from Angioproteomie and were cultured in pericyte growth medium according to manufacturer's protocols.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Permeability

Journal: Scientific Reports

Article Title: 96 perfusable blood vessels to study vascular permeability in vitro

doi: 10.1038/s41598-017-14716-y

Figure Lengend Snippet: Media optimization on microvessels using the permeability assay. ( a ) At the start of the assay (t = 0 min), the fluorescent dextrans are contained within the microvessel. Scale bar: 200 µm. ( b ) After 30 min, the differences in permeability are clearly visible: the dextran fills the gel in case of a completely permeable microvessel, while an impermeable vessel retains the dextran within the lumen of the vessel. The dotted line indicates the position of the cell barrier. Scale bar: 200 µm. ( c ) Visualization of dextran diffusion after 30 minutes for an array of 86 microvessels with three different culture conditions (M199 medium = M199, MV2 medium = MV2, Pericyte medium = PC). Excluded vessels due improper barrier formation or gel seeding are indicated with a diagonal hatch pattern. Scale bar: 2 mm. ( d ) Quantification of the permeability over different days for M199 (n = 28), MV2 (n = 29) and PC (n = 29). Data is presented as mean ± SD.

Article Snippet: MV2 endothelial cell Growth Medium (Promocell, C-22022) and Pericyte Growth Medium SR Formulation (Angioproteomie, cAP-09B) were supplemented with 1% pen/strep (Sigma, P4333).

Techniques: Permeability, Diffusion-based Assay