Journal: Heliyon

Article Title: Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization

doi: 10.1016/j.heliyon.2019.e01978

Figure Lengend Snippet: Cell Viability of (a). HUVEC (b). pericytes and (c). MDA-MB 231 cells when Treated with Propolis Extracts. The cells were treated with various concentrations of D01 and D02. After 48 hours of incubation, 10μl/well of the CCK-8 solution was added and the absorbance at 450nM was measured. Data are expressed as absorbance values with normalization to the basal values of cells with DMSO as the control, which show means ±SD and represent biological replicates. Taxol was used as a positive control of anti-angiogenesis to be compared with the samples. The best fit curve was produced by analyzing the predicted residual error sum of squares (PRESS). Bullets signify the actual average experimental data. The experiment was repeated three times with five technical replicates in each experiment. Data of D02 did not produce a converged curve on HUVECs' and pericytes' curves. Furthermore, D02 barely exhibited an inhibitory or cytotoxic effect on MDA-MB-231 cells (the plot is not depicted for efficiency).

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and placenta-derived pericytes were from Promocell, Heidelberg, Germany.

Techniques: Incubation, CCK-8 Assay, Positive Control, Produced

Journal: Heliyon

Article Title: Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization

doi: 10.1016/j.heliyon.2019.e01978

Figure Lengend Snippet: IC 50 of D01 and pure compounds on (a). HUVECs (b). pericytes and (c). MDA-MB-231 cells (n = 3). The graphs of CAPE and pinocembrin are not shown.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and placenta-derived pericytes were from Promocell, Heidelberg, Germany.

Techniques: Standard Deviation

Journal: Heliyon

Article Title: Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization

doi: 10.1016/j.heliyon.2019.e01978

Figure Lengend Snippet: Wound Healing/Cell Migration Rate of (a). HUVECs (b). pericytes and (c). MDA-MB-231 cells when Exposed to Propolis Samples for 8 Hours. In 24 well-plates, the cells were grown until confluent before wounding by a mechanical wounder to produce 11 lesions on the cell monolayers. 10 μg/ml final concentration of mitomycin-C was added 2 hours prior to wounding. Next, the cells were treated with D01 and D02 at the concentrations where they do not induce cytotoxicity. DMSO 0.1% was used as the vehicle control. After 8 hours, all wells were fixed using methanol and the resulted images were analyzed by ImageJ. Data are expressed as average % recovery values compared to the time-zero control with no normalization and represent biological replicates. An ordinary one-way ANOVA was performed with a Dunnett's multiple comparisons test where the mean of each group was compared with the mean of the vehicle control group for each drug separately. Significance is reported as a * which represents a recovery effect (* for P < 0.05, ** for P < 0.01, and *** for P < 0.001) and a # which represents an inhibitory effect (# for P < 0.05, ## for P < 0.01, and ### for P < 0.001). The experiment was repeated two times with four technical replicates in each experiment. Figure (d) is the example of photomicrograph of wound healing experiment using D01 on MDA-MB-231 cells.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and placenta-derived pericytes were from Promocell, Heidelberg, Germany.

Techniques: Migration, Concentration Assay

Journal: Heliyon

Article Title: Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization

doi: 10.1016/j.heliyon.2019.e01978

Figure Lengend Snippet: Effect of Propolis Samples on CoCl 2 -induced Pericyte Loss. Pericytes were treated with various concentrations of (a) D01 and (b) D02. After 2 hours, each well, except the control wells, were further added with CoCl 2, giving the final concentration of 250μM. After 24 hours of incubation, 10μl/well of CCK-8 solution was added and the absorbances of 450nM was measured. Data are expressed as absorbance values with normalization to the basal values of cells with DMSO as the control, which show means ±SD and represent biological replicates. An ordinary one-way ANOVA was performed with a Tukey's multiple comparisons test. The DMSO + CoCl 2 group was compared to the vehicle control group (DMSO), and the other treatments were compared to the DMSO + CoCl 2 group in order to assess the effect of propolis treatment against pericyte dropout, either stimulatory/recovery (*) or inhibitory/cytotoxic (#). Significance is reported as a * which represents a stimulatory effect (* for P < 0.05, ** for P < 0.01, and *** for P < 0.001) and # which represents an inhibitory or cytotoxic effect (# for P < 0.05, ## for P < 0.01, and ### for P < 0.001). While in (c), the protection percentage of propolis-samples-treated pericytes after experiencing a CoCl 2 -induced decline (pericyte loss) is depicted. Data were obtained by calculating the percentage of change between the data of each sample to the control data (samples which were treated by CoCl 2 only). The experiment was repeated two times with five technical replicates in each experiment.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and placenta-derived pericytes were from Promocell, Heidelberg, Germany.

Techniques: Concentration Assay, Incubation, CCK-8 Assay

Journal: Heliyon

Article Title: Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization

doi: 10.1016/j.heliyon.2019.e01978

Figure Lengend Snippet: Photomicrographs of pericytes after the incubation with D01 for 48 hours. Pericytes were treated with various concentrations of D01: (a) 1000 μg/ml, (b) 100 μg/ml, (c) 10 μg/ml, (d) 1 μg/ml, (e) DMSO, and (f) 8nM Taxol. The images were taken after 48 hours of incubation and before addition of CCK-8 solution using Nikon Phase Contrast ELWD 0.3 microscope at 80X magnification.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and placenta-derived pericytes were from Promocell, Heidelberg, Germany.

Techniques: Incubation, CCK-8 Assay, Microscopy

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Cell viability of control pericytes ( A ) and pericytes treated with AuNPs at 10 ( B ), 20 ( C ), 30 ( D ), 40 ( E ), and 50 ppm ( F ) concentrations. The graph shows the quantitative data for pericyte viability ( G ). *** p < 0.001. Bar: 100 μm.

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Pericyte proliferation and PDGFR-β mRNA expression after treatment with 30 ppm AuNPs for 24 h. Representative images are shown for the control group ( A ) and the 30 ppm AuNP-treated pericyte group ( B ). The graphs show the Ki-67 ( C ) and PDGFR-β ( D ) mRNA expression levels. *** p < 0.001. Bar: 100 nm.

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques: Expressing

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Cell migration of pericytes after treatment with 30 ppm AuNPs. Representative images are shown for the control group ( A , C ) magnified view of ( A ) and the 30 ppm AuNP-treated pericyte group ( B , D ) magnified view of ( B ). The graph shows the numbers of migrating cells ( E ). *** p < 0.001. Bars: 100 μm.

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques: Migration

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Transmission electron microscopy of pericytes in the control group at low ( A ) and high magnifications ( B ). The pericytes showed normal ultrastructure. Note the healthy mitochondria (arrowhead). Bars: 5000 nm ( A ), 1000 nm ( B ).

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques: Transmission Assay, Electron Microscopy

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Transmission electron microscopy of the 30 ppm AuNP-treated pericyte group at low ( A ) and high magnifications ( B – F ). Representative images are shown for late endosomes containing AuNPs and multivesicular bodies ( B ), autolysosomes containing only AuNPs ( C ), and autolysosomes containing AuNPs and intracellular debris ( D ). Affected mitochondria showed mild to severe swelling ( E , F ). Note the AuNPs (electron-dense dots), multivesicular bodies (black arrows), intracellular debris (double black arrows), unaffected mitochondria (black arrowhead), and swollen or damaged mitochondria (white arrowhead). Bars: 1000 nm ( A ), 100 nm ( B – D ), 500 nm ( E , F ).

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques: Transmission Assay, Electron Microscopy

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Tube formation of the control group (in which intact endothelial cells were cocultured with intact pericytes) ( A ) and the AuNP-treated group (in which intact endothelial cells and pericytes were pretreated with AuNPs) ( B ). The graph shows the numbers of tubes ( C ). Note the cluster of mixed pericytes and endothelial cells (arrow), the thick wall of tube formation (arrowhead), and the area of incomplete tubes (asterisk). *** p < 0.001. Bar: 500 µm.

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Gold Nanoparticles Affect Pericyte Biology and Capillary Tube Formation

doi: 10.3390/pharmaceutics13050738

Figure Lengend Snippet: Immunofluorescence staining of pericytes during tube formation in the control group ( A , B ) and the 30 ppm AuNP-pretreated pericyte group ( C , D ). PDGFR-β-immunopositive cells are pericytes (green pseudocolor), and unstained cells are endothelial cells. In the treated group, pericytes were round and did not extend processes. Note that the endothelial cells and AuNP-treated pericytes formed thicker walls than the control cells. The nuclei are stained with DAPI (blue pseudocolor). Bars: 100 µm.

Article Snippet: Human placental pericytes (hPC-PLs; Cat. no. C-12980, PromoCell, Heidelberg, Germany) were cultured in pericyte growth medium (Cat. no. C-28040, PromoCell, Heidelberg, Germany) supplemented with 10% SupplementMix (Cat. no. C-39840, PromoCell, Heidelberg, Germany) and 1% antibiotic-antimycotic solution (Cat. no. 15240062, Gibco, New York, NY, USA).

Techniques: Immunofluorescence, Staining

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

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:

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

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, Plasmid Preparation, Concentration Assay

Journal: Stem Cells Translational Medicine

Article Title: Perivascular Progenitor Cells Derived From Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy

doi: 10.5966/sctm.2015-0342

Figure Lengend Snippet: CD140B+CD44+ cells are derived from human embryonic stem cells (hESCs) in a matrix-dependent manner. (A): Schematic representation of matrix-dependent differentiation process for CD140B+CD44+ cells. (B): Two-color flow cytometry was used to check mesodermal specification following the differentiation schedule. All single cells were labeled with antibodies against CD140B and CD44. Obtained cells are distinguished from CD140B− and CD140B+ by the horizontal lines in each plot. Cases #1 and #2 are typical representative aspects, depending on a different starting H9-hESC line. After collagen-dependent natural selection through a simple medium change, the attached CD140B+ population showed a strong CD44 expression at Day 7 (A). Values in each quadrant plot represent percentage of population, mean ±SD (n = 6). (C, D): The change of gene context by collagen-dependent attachment step showed a big difference. Differential gene expression profiling was performed among H9-hESCs, embryoid bodies (EBs) induced by bone morphogenetic protein 4 (BMP4) (EBs) and naturally selected CD140B+CD44+ population at Day 7 (perivascular progenitor cells, PVPCs, CD140B+CD44+ population) from three independent batches. (C): Heatmap of each group. Each column represents a single microarray analysis. (D): Hierarchical clustering analysis of the global gene expression proϕiles using the average linkage and the Pearson distance. (E): Expanded CD140B+CD44+ population exhibited a unique cell morphology. Scale bar = 20 μm. (F): The cell proliferation was monitored with the CCK-8 assay at different time points. Data are means of three separated experiments ± SD. Abbreviations: BM-MSC, human bone marrow-derived mesenchyme stem cells; h, hour; hPL-PC, human placenta-derived pericyte; MEF, mouse embryonic fibroblast; OD, optical density.

Article Snippet: Both human placenta-derived pericytes (hPL-PCs) and human bone marrow-derived MSCs (BM-MSCs) were obtained from PromoCell (Heidelberg, Germany, http://www.promocell.com/ ).

Techniques: Derivative Assay, Flow Cytometry, Labeling, Selection, Expressing, Microarray, CCK-8 Assay

Journal: Stem Cells Translational Medicine

Article Title: Perivascular Progenitor Cells Derived From Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy

doi: 10.5966/sctm.2015-0342

Figure Lengend Snippet: Naturally selected CD140B+CD44+ cells show the characteristics of multipotent perivascular progenitor cells (PVPCs). (A): Expanded CD140B+CD44+ cells (human embryonic stem cells derived from PVPCs; hESC-PVPCs) were labeled with the indicated antibodies. The overlaid open histogram displays immunoglobulin control against each target. Values in each quadrant plot represent percentage of population, mean ± SD (n = 8). (B): Generalized pericyte marker, NG2 (green) expressed in both hESC-PVPCs and human primary pericytes (hPL-PC). Nuclear DNA was stained with 4′,6-diamidino-2-phenylindole (blue). (C): Skeletogenic differentiation potential of hESC-PVPCs was shown in the differentiation condition of adipogenesis (left, Oil red staining) and osteogenesis (right, Alizarin red staining). (D): Smooth muscle cell (SMC) differentiation potential of hESC-PVPCs was shown in SMC differentiation media (SMDM). Quantitative polymerase chain reaction values represent mean (n = 3) ± SD (∗∗∗, p < .0001). SMC-specific markers, α-SMA (green), and CNN (red) were detected in hESC-PVPC after 6 days in SMDM differentiation. (E): Dye transfer (circle in the plot, yellow-green) increased in a coculture of DiI-labeled hESC-PVPC (PVPC-DiI, red) and Calcein-labeled other vascular lineages (upper, human umbilical vein endothelial cell, HUVEC-Calcein; lower, umbilical artery smooth muscle cell, UASMC-Calcein, green). Values in each plot represent percentage of population, mean ± SD (n = 3). Dye-transferred hESC-PVPCs were also detected on culture well (white dots). (F, G): Perivascular localization of hESC-PVPC (PVPC, DiI, red) was confirmed in three-dimensional fibrin gel bead assay with HUVEC (DiO, green). Magnified view (G) of rectangular region (F), and orthogonal projection images (white dotted lines 1 and 2). All scale bars = 100 μm. Abbreviations: α-SMA, α-smooth muscle actin; CNN, Calponin 1; d, days.

Article Snippet: Both human placenta-derived pericytes (hPL-PCs) and human bone marrow-derived MSCs (BM-MSCs) were obtained from PromoCell (Heidelberg, Germany, http://www.promocell.com/ ).

Techniques: Derivative Assay, Labeling, Marker, Staining, Real-time Polymerase Chain Reaction