primary human placental pericytes hpc pl  (PromoCell)

Journal: Lab on a chip

Article Title: Late-stage placental barrier model for transport studies of prescription drugs during pregnancy.

doi: 10.1039/d5lc00075k

Figure Lengend Snippet: Fig. 3 Effects of pericytes on vascular and total permeability of placental barrier model on days 16 and 18 of culture. A. Brightfield images of vascular channel for the following conditions: no cell control, ST only, ST + HUVECs, and two tricultures with ST + HUVECs + hPC-PLs at high and low density. Fluorescent images of 4 kDa (green) and 65 kDa (red) dextran permeation through the endothelial channel after 1 hour of perfusion. B. Vascular barrier permeability to 4 kDa dextran. (N = 3–7, one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001) C. Vascular barrier permeability to 65 kDa dextran. (N = 3–7, one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001) D. Fluorescent images of 65 kDa dextran (red) permeating from the central maternal compartment, through the ST and HUVEC barriers, to the adjacent fetal compartments after 24 hours. E. Total barrier permeability to 4 kDa dextran from the maternal to the fetal compartments. (N = 6–7, one-way ANOVA, **p < 0.01, ***p < 0.001) F. Total barrier permeability to 65 kDa dextran from the maternal to the fetal compartments. (N = 6–7, one-way ANOVA, ***p < 0.001).

Article Snippet: Primary human placental pericytes (hPC-PL) were obtained from PromoCell (C-12980), cultured in pericyte growth medium 2 (PGM2, Millipore Sigma, C-28041) and used until passage 5.

Techniques: Permeability, Control

Journal: Placenta

Article Title: Characterizing placental pericytes: Hypoxia and proangiogenic signalling.

doi: 10.1016/j.placenta.2024.07.314

Figure Lengend Snippet: Fig. 1. Human placental pericyte cell count and viability in 20 % O2 and 1 % O2 (n = 3 per treatment group). Placental pericyte cell count (A) and viability (B) were unchanged in 1 % O2. Statistical analysis compared cell count/viability in 20 % O2 and 1 % O2 using unpaired t-tests. Data expressed as mean ± SEM. Stars indicate statistical significance between treatment groups, *p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001.

Article Snippet: Primary human placental pericytes (PromoCell) were seeded at 3.0 × 103 cells/cm2 in Basal Medium (PromoCell) supplemented with supplier-provided SupplementMix and used up to passage 6 (recommended by supplier).

Techniques: Cell Counting

Journal: Placenta

Article Title: Characterizing placental pericytes: Hypoxia and proangiogenic signalling.

doi: 10.1016/j.placenta.2024.07.314

Figure Lengend Snippet: Fig. 2. Angiogenic secretion of human placental pericytes in 20 % O2 and 1 % O2 (n = 3 per treatment group). ELISAs performed on cell culture supernatant. (A) Placental pericyte secretion of ANG1 was decreased in 1 % O2. (B) Placental pericyte secretion of VEGF was increased in 1 % O2. (C) Placental pericyte secretion of MMP2 was unchanged in 1 % O2. Statistical analysis compared secreted angiogenic factor concentration in 20 % O2 and 1 % O2 using unpaired t-tests. Data expressed as mean ± SEM. Stars indicate statistical significance between treatment groups, *p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001.

Article Snippet: Primary human placental pericytes (PromoCell) were seeded at 3.0 × 103 cells/cm2 in Basal Medium (PromoCell) supplemented with supplier-provided SupplementMix and used up to passage 6 (recommended by supplier).

Techniques: Cell Culture, Concentration Assay

Journal: Placenta

Article Title: Characterizing placental pericytes: Hypoxia and proangiogenic signalling.

doi: 10.1016/j.placenta.2024.07.314

Figure Lengend Snippet: Fig. 5. PlGF and sFLT1 secretion of human placental pericytes, HUVECs, BeWos (cytotrophoblasts), and syncytialized BeWos in 20 % O2 and 1 % O2 (n = 3 per treatment group). ELISAs performed on cell culture supernatant. (A) HUVEC and syncytialized BeWo secretion of PlGF were increased in 1 % O2. (B) HUVEC, BeWo (cytotrophoblast), and syncytialized BeWo secretion of sFLT1 were increased in 1 % O2, while placental pericyte secretion of sFLT1 was decreased in 1 % O2. Statistical analysis compared secreted angiogenic factor concentration in 20 % O2 and 1 % O2 using unpaired t-tests for each cell type. Data expressed as mean ± SEM. Stars indicate statistical significance between treatment groups, *p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001.

Article Snippet: Primary human placental pericytes (PromoCell) were seeded at 3.0 × 103 cells/cm2 in Basal Medium (PromoCell) supplemented with supplier-provided SupplementMix and used up to passage 6 (recommended by supplier).

Techniques: Cell Culture, Concentration Assay

Journal: Placenta

Article Title: Characterizing placental pericytes: Hypoxia and proangiogenic signalling.

doi: 10.1016/j.placenta.2024.07.314

Figure Lengend Snippet: Fig. 6. Double IF on HUVEC- and human placental pericyte-derived vascular sprout. 200× magnification of HUVEC and human placental pericyte-coated bead and vascular sprout stained for nuclei (A, NucBlue, blue), endothelial cells (B, VE cadherin, red) and pericyte cells (C, αSMA, green). Human placental pericytes colocalize with HUVECS on the endothelial sprouts in vitro (D, merge). Scale bar = 150 μm.

Article Snippet: Primary human placental pericytes (PromoCell) were seeded at 3.0 × 103 cells/cm2 in Basal Medium (PromoCell) supplemented with supplier-provided SupplementMix and used up to passage 6 (recommended by supplier).

Techniques: Derivative Assay, Staining, In Vitro

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA actively downregulates TF in primary human pericytes. Pericyte cultures were treated with PMA for the indicated times. The mean +/− standard deviation (SD) of 3 independent experiments is shown. *p<0.05, **p<0.01, ***p<0.001. A. Expression of TF protein was assessed by western blot. B. For each time point the relative amount of TF protein expressed by PMA-treated cells is shown as a percentage of TF expressed by vehicle-treated cells. C. Activity of total TF as measured in lysed pericytes was determined by measuring cleavage of a FXa-specific chromogenic substrate as described in “Methods”. TF activity is presented as the rate of FXa generation. D. Expression of TF mRNA was quantitated by qRT-PCR. For each time point the amount of TF mRNA expressed by PMA-treated cells is presented as the fold change relative to TF mRNA expressed by vehicle-treated control cells.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Standard Deviation, Expressing, Western Blot, Activity Assay, Quantitative RT-PCR, Control

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: Internalized TF is degraded primarily in lysosomes. Pericytes were treated with MG132 or chloroquine prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Relative amounts of internalized TF and expression of total TF were assessed by biotin labeling of cell surface proteins and western blotting as in “Methods”. B. The amounts of internalized TF are expressed as percentages relative to the total amount of TF expressed on the cell surface. C. Relative amounts of total TF expressed by PMA-treated pericytes are shown as percentages of TF expressed by control cells. D. Pericytes were stained for TF (green), the lysosomal marker LAMP-1 (red), and nuclei (blue, DAPI). E. Co-localization of TF and LAMP-1 was determined by calculating Pearson’s Correlation Coefficient with Image J, Fiji Plugin.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Expressing, Labeling, Western Blot, Control, Staining, Marker

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA shortens the half-life of TF protein while leaving degradation of TF mRNA unaffected. The mean +/− standard deviation (SD) of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericytes were pre-treated with Cyclohexamide (CHX) prior to addition of vehicle or PMA for the indicated times. Degradation of TF protein was assessed by western blot. B. Relative amounts of TF protein expressed by CHX-treated cells are given as a percentage of TF expressed by non-CHX-treated control cells. C. Pericytes were pre-treated with Actinomycin D (ActD) prior to receiving vehicle or PMA for the indicated times. Degradation of TF mRNA was analyzed by qRT-PCR. The amount of TF mRNA expressed by ActD-treated pericytes relative to non-ActD-treated control cells is shown.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Standard Deviation, Western Blot, Control, Quantitative RT-PCR

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: Inhibition of Protein Kinase C attenuates PMA-mediated downregulation of TF. Pericytes were pre-treated with Go 6983 or GFX prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. *p<0.5, **p<0.1, ***p<0.001. A. TF protein expression was analyzed by western blot. B. For each time point the amount of TF protein is shown as a percentage of the TF expressed by control cells. C. Expression of TF mRNA was quantified by qRT-PCR. The amount of expressed TF mRNA is presented as the fold change relative to TF mRNA expressed by control cells. D. Activity of total TF in lysed pericytes is shown as the rate of FXa generation.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Inhibition, Expressing, Western Blot, Control, Quantitative RT-PCR, Activity Assay

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA increases internalization of TF from the pericyte surface. Where specified, pericytes were pre-treated with either Go 6983 or GFX prior to receiving PMA for the times indicated. The mean +SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericyte cultures were fixed and stained for TF (green). Prominent punctate staining for TF was seen 4 hours after PMA while TF was virtually absent 8 hours after PMA. B. The total TF content of pericytes was determined by western blotting of whole cell lysates. Internalized surface TF was determined by labeling cell surface proteins with biotin, isolating biotin-labeled proteins from cell lysates, and assessing their TF content by western blotting. C. Relative amounts of internalized TF are shown as a percentage of the total surface TF. D. Relative amounts of total TF are presented as a percentage of the total surface TF expressed by vehicle-treated control cells. E. Activity of surface TF was determined on whole cells by measuring the rate of FXa generation.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Staining, Western Blot, Labeling, Control, Activity Assay