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: BMC Ophthalmology

Article Title: Anti-angiogenic properties of rapamycin on human retinal pericytes in an in vitro model of neovascular AMD via inhibition of the mTOR pathway

doi: 10.1186/s12886-022-02334-w

Figure Lengend Snippet: Toxicity of Rapamycin on HUVEC ( A ) and Human retinal pericytes ( B ). No toxicity on both HRP and HUVEC could be detected within the whole range of solubility

Article Snippet: The desired concentration of rapamycin was achieved through dilution in ready-to-use pericyte (Cell Systems, Seattle, USA) or endothelial cell growth medium (PromoCell, Heidelberg, Germany) with a final concentration of 0.1% DMSO for in vitro studies.

Techniques: Solubility

Journal: BMC Ophthalmology

Article Title: Anti-angiogenic properties of rapamycin on human retinal pericytes in an in vitro model of neovascular AMD via inhibition of the mTOR pathway

doi: 10.1186/s12886-022-02334-w

Figure Lengend Snippet: Effects of Rapamycin on proliferation ( A ) and migration ( B ) in pericytes A. Rapamycin significantly reduced proliferation in pericytes starting from 0.005μg/ml with a half maximal inhibitory concentration of 423 ng/ml. B. For all concentrations tested, Rapamycin dose-dependently reduced pericyte migration into the scratch wound after 24 hours (Co – Rapa 0.005: p =0.006; MD: 25.7; 95 % CI: 9.2 – 42.2; Co – Rapa 0.05: p <0.0001; MD 42.0; 95 % CI: 25.5 – 58.4; Co – Rapa 0.5: p <0.0001; MD 56.7; 95 % CI: 40.2 – 73.2; Co – Rapa 5: p <0.0001; MD 76.8; 95 % CI: 60.3-93.3). The control wound was densely covered with HRP after 24 hours in contrast to the highest used concentration of 5μg/ml where only little migration was present ( right )

Article Snippet: The desired concentration of rapamycin was achieved through dilution in ready-to-use pericyte (Cell Systems, Seattle, USA) or endothelial cell growth medium (PromoCell, Heidelberg, Germany) with a final concentration of 0.1% DMSO for in vitro studies.

Techniques: Migration, Concentration Assay

Journal: BMC Ophthalmology

Article Title: Anti-angiogenic properties of rapamycin on human retinal pericytes in an in vitro model of neovascular AMD via inhibition of the mTOR pathway

doi: 10.1186/s12886-022-02334-w

Figure Lengend Snippet: Dose dependent effects of rapamycin on pericyte migration

Article Snippet: The desired concentration of rapamycin was achieved through dilution in ready-to-use pericyte (Cell Systems, Seattle, USA) or endothelial cell growth medium (PromoCell, Heidelberg, Germany) with a final concentration of 0.1% DMSO for in vitro studies.

Techniques:

Journal: BMC Ophthalmology

Article Title: Anti-angiogenic properties of rapamycin on human retinal pericytes in an in vitro model of neovascular AMD via inhibition of the mTOR pathway

doi: 10.1186/s12886-022-02334-w

Figure Lengend Snippet: Effects of rapamycin on pericyte migration compared to untreated control

Article Snippet: The desired concentration of rapamycin was achieved through dilution in ready-to-use pericyte (Cell Systems, Seattle, USA) or endothelial cell growth medium (PromoCell, Heidelberg, Germany) with a final concentration of 0.1% DMSO for in vitro studies.

Techniques: Migration

Journal: BMC Ophthalmology

Article Title: Anti-angiogenic properties of rapamycin on human retinal pericytes in an in vitro model of neovascular AMD via inhibition of the mTOR pathway

doi: 10.1186/s12886-022-02334-w

Figure Lengend Snippet: Effects of Rapamycin on tube formation in a HUVEC monoculture ( A ) compared to a HUVEC/HRP co-culture ( B ). Tube formation after 6 hours is significantly reduced under Rapamycin compared to the untreated control leading to a decrease down to 74.0%, 63.6%, 59.1% and 54.9% respectively (Co – Rapa 0.005: p =0.001; MD: 26.0; 95 % CI: 13.8 – 38.2; Co – Rapa 0.05: p <0.0001; MD 36.4; 95 % CI: 24.2 – 48.6; Co – Rapa 0.5: p <0.0001; MD 40.9; 95 % CI: 28.7 – 53.1; Co – Rapa 5: p <0.0001; MD 45.1; 95 % CI: 33.0-57.3). However, HUVEC monoculture tubes destabilize also without treatment after 30 hours while still being steady after the same time frame by adding pericytes and thus creating a co-culture comparable to the in vivo condition. B: Adding Rapamycin to the co culture tubes seems to disrupt endothelial tube stabilization (Co – Rapa 0.005: p <0.0001; MD: 45.1; 95 % CI: 30.2 – 59.9; Co – Rapa 0.05: p <0.0001; MD 78.9; 95 % CI: 64.1 – 93.8; Co – Rapa 0.5: p <0.0001; MD 80.0; 95 % CI: 65.1 – 94.8; Co – Rapa 5: p <0.0001; MD 94.0; 95 % CI: 79.1-108.9)

Article Snippet: The desired concentration of rapamycin was achieved through dilution in ready-to-use pericyte (Cell Systems, Seattle, USA) or endothelial cell growth medium (PromoCell, Heidelberg, Germany) with a final concentration of 0.1% DMSO for in vitro studies.

Techniques: Co-Culture Assay, In Vivo