pericyte growth media  (PromoCell)

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: TGF β -1 signalling via canonical receptor ALK5 in placental pericytes. ( a ) Representative image of ALK5/TGF β R1 expression (ICC/IF; green), WGA plasma membrane staining (red), nuclear stain (blue). Scale bar 150 μm; ( b ) RT-PCR analysis of Type I ALK5 TGFBR1 (91 bp) and Type II receptor, TGFBR2 (99 bp) in placental pericytes (n = 3); ( c ) pSMAD2 (52kD) and GAPDH (37kD) Western Blot. Treatment groups (n = 3): VEH control (DMSO), TGF β -1 (10 ng/mL), TGF β -1 + ALK5 in. (10 ng/mL + 2 μM, respectively); ( d ) Western Blot densitometry analysis; one-way ANOVA. Stars indicate statistical significance between treatment groups, ns = no significance, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. Error bars (SEM).

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Expressing, Clinical Proteomics, Membrane, Staining, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: ELISA analysis of angiogenic factors secreted from placental pericytes. Treatment groups (n = 3): VEH control (DMSO), TGF β -1 (10 ng/mL), TGF β -1 + ALK5 in; ( a ) VEGF secretion (Kruskal–Wallis analysis); ( b ) MMP-2 secretion (one-way ANOVA). ( c ) ANG-1 secretion (one-way ANOVA). ELISA results normalized to cell number. Stars indicate statistical significance between treatment groups, ns = no significance, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. Error bars (SEM).

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: Collagen IV, fibronectin and laminin staining in placental pericytes. Representative ICC/IF images (VEH-treated placental pericytes); ( a ) Collagen IV (red), nuclear stain (blue); data expressed as area and mean fluorescent intensity (MFI); one-way ANOVA, normalized to cell number; ( b ) Fibronectin (red), nuclear stain (blue), data expressed as area and MFI; one-way ANOVA, normalized to cell number; ( c ) Laminin (red), nuclear stain (blue); data expressed as area; Kruskal–Wallis analysis and MFI; one-way ANOVA, normalized to cell number. Scale Bars = 150 μm. Stars indicate statistical significance between treatment groups, ns = no significance, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. Error bars (SEM).

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Staining

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: Placental pericyte secretion of inflammatory factors. Analysis of inflammatory secretion (ELISA). Treatment groups: VEH control (DMSO), TGF β -1 (10 ng/mL), TGF β -1 + ALK5 in. (10 ng/mL + 2 μM, respectively); ( a ) IL-6 secretion; one-way ANOVA; ( b ) MCP-1 secretion; one-way ANOVA; ( c ) sVCAM-1 secretion; one-way ANOVA; ( d ) CX3CL1 secretion; one-way ANOVA. ( e ) IL-8 secretion; Kruskal–Wallis analysis. Data (n = 3); stars indicate statistical significance between treatment groups ns = no significance, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. Error bars (SEM).

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: Alternate TGF β -1 signalling pathway in placental pericytes: Type I receptor ACVRL1 (ALK1, 94 bp) expression in placental pericytes, as assessed by RT-PCR; original image shown.

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: The regulation of placental pericyte function through transforming growth factor β -1 signalling

doi: 10.1038/s41598-025-20432-9

Figure Lengend Snippet: Phagocytosis capacity of placental pericytes. Treatment groups: VEH control and TGF β -1 (10 ng/mL) with and without FluoSphere incubation; ( a ) Representative flow cytometry histograms to identify the percentage of the population with and without phagocytosed FluoSpheres based on fluorescence; ( b ) Analysis of phagocytosis (Mann–Whitney). Data (n = 3) expressed as mean ± SEM. Stars indicate statistical significance, ns = no significance, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; c Fluorescent and phase contrast images of pericyte phagocytosis, nuclei (blue), FluoSphere (green). Yellow arrows identify pericytes with phagocytosed beads, and red arrows identify beads that have not been phagocytosed. Scale bar = 150 μm.

Article Snippet: To assess pericyte phagocytosis , FACS-isolated primary human placental pericytes (PromoCell) were cultured in ready-to-use pericyte growth media (PromoCell), seeded at 3.0 × 10 3 cells/cm 2 and allowed to establish in 5% CO 2 in 37 °C humidified air for 48 h before TGF β -1 treatment.

Techniques: Control, Incubation, Flow Cytometry, Fluorescence, MANN-WHITNEY

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: Cellular components of BBB are productively infected with icSARS-CoV-2-nMG. ( A – C ) Live-cell imaging of HUVECs, astrocytes, and pericytes mock infected or infected with 1 MOI of icSARS-CoV-2-nMG at 24 hours post infection (hpi). Brightfield and green fluorescent images are merged to demonstrate infection within cells. Yellow arrows indicate infected cells. ( D – F ) Representative plaque assay images. Numbers indicate dilution of cell culture supernatant from virus-infected cells. Mean plaque assays conducted with 2 replicates from the supernatant of infected samples at 48 hpi. Scale bars = 400 μm. * p < 0.05 as measured using Student’s t -test.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Infection, Live Cell Imaging, Plaque Assay, Cell Culture, Virus

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: HUVECs, astrocytes, and pericytes were infected with 1 MOI of icSARS-CoV-2-nMG. Cells were fixed 48 hpi and immunostained. Cell pellets harvested at 24, 48 and 72 hpi used for qPCR analysis. ( A ) Immunostaining of infected HUVECs with ACE2 and SARS-CoV-2 spike antibodies. ( B , C ) Immunostaining of infected astrocytes and pericytes with DPP4 and SARS-CoV-2 spike antibodies. ( D – I ) qPCR analysis of N and spike mRNA as a measure of infection at 24, 48, and 72 hpi. * p < 0.05 as measured using One-Way ANOVA and subsequent post hoc Fisher’s LSD test.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Infection, Immunostaining

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: Knocking down ACE2 in HUVECs and DPP4 in astrocytes and pericytes reduces infection burden. Cells were transduced with shACE2- or shDPP4-lentiviruses or corresponding shScr lentiviruses (1 U/cell) for 48 h prior to infection with SARS-CoV-2 (1 MOI). ( A – C ) Western blots of ACE2, DPP4 and N protein expression in HUVECs astrocytes and pericytes. ( D – F ) qPCR measurements of viral mRNA for N and spike mRNAs of SARS-CoV-2 infected cells compared with their mock counterparts. ( G – I ) Representative plaques from each experimental condition for each cell type, and their quantifications in PFU/mL. * p < 0.05 using One-Way ANOVA and subsequent post hoc analysis with Fisher’s LSD Test.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Infection, Transduction, Western Blot, Expressing

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: Transwell BBB model is infected in both compartments by SARS-CoV-2 while maintaining paracellular integrity. ( A , B ) qPCR analysis of mRNA of N and spike in the upper (HUVECs) and lower wells (astrocytes and pericytes) of the model. ( C ) The permeability coefficient of passive diffusion of 3 kDa and 10 kDa fluorescent dextran molecules placed in the upper compartment of the BBB model to the lower compartment 24 hpi and 48 hpi. The raw reads of fluorescence were converted to a permeability coefficient using Equation (1) mentioned above. ( D ) Representative images of plaque assays and their corresponding dilution factor. Supernatants from the upper and lower compartments of the transwell were sampled at 48 hpi and measured in a plaque assay with infectious viral titer being reported in PFU/mL. #, * p < 0.05 using Student’s t -test or One-Way ANOVA and subsequent post hoc analysis with Fisher’s LSD Test.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Infection, Permeability, Diffusion-based Assay, Fluorescence, Plaque Assay

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: SARS-CoV-2 transcytosis across the BBB is a process mediated through clathrin and caveolin endosomes irrespective of ACE2 expression. ( A – D ) qPCR measurements of infection in the upper well quantifying N and spike mRNA expression 16 hpi demonstrating varying levels of infection in both shScr + CoV-2 and shACE2 + CoV-2. Drug conditions are compared with vehicle group. ( E – H ) Differences (or lack thereof) in paracellular permeability measured using a 3 kDa and 10 kDa fluorescent dextran molecule placed in the upper compartment of the BBB model and passive diffusion of the molecules to the lower compartment. The raw reads of fluorescence were converted to permeability coefficients using Equation (1) mentioned above. ( I – K ) Representative images of plaque assays and their corresponding dilution factor. Supernatant from the lower compartment of the transwell was sampled at 16 hpi and measured in a plaque assay with infectious viral titer being reported in PFU/mL. ( L ) Detection of N protein in the cells (astrocytes and pericytes) of the lower well using Western blots. All mock conditions demonstrate no detection, while varying levels of N were seen within the CoV-2 conditions. Detection of N protein in lower well cells was not wholly correlated with plaque assays but broadly aligns. * p < 0.05 using Two-Way ANOVA and subsequent post hoc analysis with Fisher’s LSD Test.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Expressing, Infection, Permeability, Diffusion-based Assay, Fluorescence, Plaque Assay, Western Blot

Journal: Viruses

Article Title: Modulation of Paracellular Permeability in SARS-CoV-2 Blood-to-Brain Transcytosis

doi: 10.3390/v16050785

Figure Lengend Snippet: Proposed model of SARS-CoV-2 infection and transcytosis of the BBB. ( A , B ) Direct infection of brain endothelial cells including ACE2 binding, furin priming, and S1/S2 cleavage by either TMPRSS2, or CTSB and/or furin. ( C ) Transcytosis with initial anchoring to glycocalyx, endocytosis by either caveolin or clathrin, and deposition on abluminal side of endothelium. ( D , E ) Proposed mode of infection of pericytes and astrocytes utilizing DPP4 binding, furin priming, and TMPRSS2 spike cleavage, or DPP4 binding and endocytosis for spike cleavage within endosome. ( F ) Diagram showing intact tight junctions preventing passive diffusion of SARS-CoV-2.

Article Snippet: HUVECs were cultured in a VascuLife EnGS Endothelial Medium Complete Kit (LL-0002, Frederick, MD, USA), astrocytes were grown in specialized media Astrocyte Media [#1801], and pericytes were grown in specialized media Pericyte Media [#1201] both obtained from ScienCell (Carlsbad, CA, USA).

Techniques: Infection, Binding Assay, Diffusion-based Assay

Journal: bioRxiv

Article Title: Generation of hiPSC-derived brain microvascular endothelial cells using a combination of directed differentiation and transcriptional reprogramming strategies

doi: 10.1101/2024.04.03.588012

Figure Lengend Snippet: A) Schematic showing the process of generating the 3D NVU system in the MIMETAS microfluidic system. B) Brightfield images showing HBMEC and rBMEC only tubules, as well as their co-cultures with primary astrocytes (AC) and primary pericytes (PC) or iPSC-derived astrocytes (iACs) and pericytes (iPCs), respectively. C) Representative immunofluorescence staining images for rBMECs showing endothelial marker CD31 and VE-cadherin (top panel) and junctional expression of TJ markers ZO-1 and Claudin-5 (bottom panel) in tubules formed inside 3D microfluidic channels; scale bar = 50 µm D) 3D view of representative immunofluorescence staining for MIMETAS microfluidic culture showing all three channels with PDGFRβ + pericytes and S100β + astrocytes in brain channel and rBMEC tubule in the blood channel. E) Close up view of representative immunofluorescence staining showing TJ marker ZO-1 for rBMECs, S100β for astrocytes, and PDGFRβ for pericytes in the blood channel; cross sectional view shows tubule with open lumen. F) Representative image showing differential diffusion of fluorescent tracer from leaky and tight barriers in 3D microfluidic culture. G-H) Permeability measurement of primary HBMEC tubules, HBMEC tubules with ACs and PCs, rBMEC tubules, and rBMEC tubules with iACs and iPCs in the middle ECM channel using large 70 kDa dextran and small ∼900Da Biocytin tracer over a course of 45 minutes. N=36-41 NVU chips. Data was analyzed using two-way ANOVA and presented as mean±SD for 3 independent experiments, ns, **<0.01, ****<0.0001. I) 3D TEER measurement comparing primary HBMEC tubules, HBMEC tubules with ACs and PCs, rBMEC tubules, and rBMEC tubules with iACs and iPCs; each point represents one individual NVU chip. N = 30-48. Data is presented as mean±SD, and statistical comparisons were made between each group using one-way ANOVA, ****<0.0001, *<0.05.

Article Snippet: PCs were cultured in Pericyte Media (PM, ScienCell), and ACs were cultured in Astrocyte Media (AM, ScienCell).

Techniques: Derivative Assay, Immunofluorescence, Staining, Marker, Expressing, Diffusion-based Assay, Permeability