Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Time course of NF- κ B activation in cocultured C 2 C 12 cells and pericytes. p65 DNA binding activity for cocultured C 2 C 12 cells and pericytes in uninjured control (CON) and scratch-injured (INJ) conditions at baseline (BSLN), 3, 6, and 24 h time points. Data are means ± SD. *Significantly increased compared to BSLN for C 2 C 12 cells. † Significantly increased compared to BSLN for pericytes.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Binding Assay, Activity Assay, Control

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Time course of MCP-1 secretion from cocultured pericytes and C 2 C 12 cells. Monocyte chemoattractant protein-1 (MCP-1) secretion by cocultured C 2 C 12 cells and pericytes in uninjured control (CON) and scratch-injured (INJ) conditions at baseline (BSLN), 3, 6, and 24 h time points. Data are means ± SD. *Significantly increased compared to BSLN for C 2 C 12 cells. † Significantly greater vs. C 2 C 12 cells at 24 h.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Control

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Pericyte transfection efficacy and efficiency in altering NF- κ B activation. (A) Representative images of pericytes transfected with vectors designed to enhance (constitutively active (c.a.) IKK β -EGFP), diminish (dominant negative (d.n.) IKK β -EGFP), or have no effect on (empty vector (e.v.) pEF6/HisB) NF- κ B activity at 24 h post transfection. (B) A luciferase reporter system was used to assess the ability of expression plasmids to alter NF- κ B expression. Data are means ± SD.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Transfection, Activation Assay, Dominant Negative Mutation, Plasmid Preparation, Activity Assay, Luciferase, Expressing

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: HMVEC proliferation is affected by pericyte NF- κ B activation in coculture. Human microvascular endothelial cell (HMVEC) number in coculture with pericytes expressing constitutively active IKK β (c.a.), dominant negative IKK β (d.n.), or empty vector control (e.v.). Data are means ± SD. *Significant difference between c.a. and d.n.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Expressing, Dominant Negative Mutation, Plasmid Preparation, Control

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Pericyte NF- κ B activation affects cytokine secretion in pericyte/HMVEC cocultures. Cytokine secretion of granulocyte-colony stimulating factor (G-CSF), fractalkine, interleukin 6 (IL-6), interleukin 7 (IL-7), interleukin 8 (IL-8), interferon gamma-induced protein-10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted (RANTES), and eotaxin in the cocultures of human microvascular endothelial cells (HMVECs) and pericytes that were genetically altered to express constitutively active IKK β (c.a.) or dominant negative IKK β (d.n.) in comparison to empty vector control (e.v.). Significantly greater cytokine concentration was observed in the c.a. IKK β condition compared to both e.v. and d.n. IKK β conditions for all cytokines ( P < 0.05).

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Dominant Negative Mutation, Comparison, Plasmid Preparation, Control, Concentration Assay

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