pbmc isolation peripheral blood mononuclear cells pbmcs  (Mediatech)

 
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    Structured Review

    Mediatech pbmc isolation peripheral blood mononuclear cells pbmcs
    S1P modulation reduces transmigration of <t>PBMCs</t> across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in <t>PBMC</t> migration after 1 hour compared to T I. *p
    Pbmc Isolation Peripheral Blood Mononuclear Cells Pbmcs, supplied by Mediatech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc isolation peripheral blood mononuclear cells pbmcs/product/Mediatech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmc isolation peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-04
    86/100 stars

    Images

    1) Product Images from "Sphingosine 1 Phosphate at the Blood Brain Barrier: Can the Modulation of S1P Receptor 1 Influence the Response of Endothelial Cells and Astrocytes to Inflammatory Stimuli?"

    Article Title: Sphingosine 1 Phosphate at the Blood Brain Barrier: Can the Modulation of S1P Receptor 1 Influence the Response of Endothelial Cells and Astrocytes to Inflammatory Stimuli?

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0133392

    S1P modulation reduces transmigration of PBMCs across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in PBMC migration after 1 hour compared to T I. *p
    Figure Legend Snippet: S1P modulation reduces transmigration of PBMCs across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in PBMC migration after 1 hour compared to T I. *p

    Techniques Used: Transmigration Assay, In Vitro, Co-Culture Assay, Migration

    2) Product Images from "IL-6 blockade suppresses the blood-brain barrier disorder, leading to prevention of onset of NMOSD"

    Article Title: IL-6 blockade suppresses the blood-brain barrier disorder, leading to prevention of onset of NMOSD

    Journal: bioRxiv

    doi: 10.1101/2021.01.28.428564

    Construction of ex-vivo BBB model with triple co-culture system for leukocyte transmigration and effect of satralizumab on NMO-IgG-induced transmigration of leukocytes in ex-vivo . A) 3D flow chamber and 3D flow membrane. the triple-cultured membranes were transferred in 3D chamber. B) Whole set up of migration assay under shear forces. Normal human peripheral blood mononuclear cells (PBMC) flowed onto luminal side with physiological shear force by peristatic pomp. C) Schema of 3D flow chamber in transmigration assay. Total migrated cells were recovered from the bottom chamber and enumerated. D) Confocal 3D analysis with living staining of BBB cell lines and PBMCs in transmigration assay. PBMC were stained with Cell trackerGreen and all human BBB cell lines (hECs, hPCTs, and hASTs) were stained with CytoID Red. Some PBMCs protruded through the membrane pores. E) Schema of leukocyte transmigration assay with NMO-IgG and/or Satralizumab. F) Flow-based leukocyte transmigration assays utilizing a 3D flow Chamber showed that application of NMO-IgG increased the numbers of total migrating PBMCs and CD4 + , CD8 + , and CD19 + cells relative to numbers migrating with Control IgG, and that the application of NMO-IgG plus satralizumab significantly suppressed that increase. *P
    Figure Legend Snippet: Construction of ex-vivo BBB model with triple co-culture system for leukocyte transmigration and effect of satralizumab on NMO-IgG-induced transmigration of leukocytes in ex-vivo . A) 3D flow chamber and 3D flow membrane. the triple-cultured membranes were transferred in 3D chamber. B) Whole set up of migration assay under shear forces. Normal human peripheral blood mononuclear cells (PBMC) flowed onto luminal side with physiological shear force by peristatic pomp. C) Schema of 3D flow chamber in transmigration assay. Total migrated cells were recovered from the bottom chamber and enumerated. D) Confocal 3D analysis with living staining of BBB cell lines and PBMCs in transmigration assay. PBMC were stained with Cell trackerGreen and all human BBB cell lines (hECs, hPCTs, and hASTs) were stained with CytoID Red. Some PBMCs protruded through the membrane pores. E) Schema of leukocyte transmigration assay with NMO-IgG and/or Satralizumab. F) Flow-based leukocyte transmigration assays utilizing a 3D flow Chamber showed that application of NMO-IgG increased the numbers of total migrating PBMCs and CD4 + , CD8 + , and CD19 + cells relative to numbers migrating with Control IgG, and that the application of NMO-IgG plus satralizumab significantly suppressed that increase. *P

    Techniques Used: Ex Vivo, Co-Culture Assay, Transmigration Assay, Cell Culture, Migration, Staining, Leukocyte Transmigration Assay

    Related Articles

    Isolation:

    Article Title: Sphingosine 1 Phosphate at the Blood Brain Barrier: Can the Modulation of S1P Receptor 1 Influence the Response of Endothelial Cells and Astrocytes to Inflammatory Stimuli?
    Article Snippet: F-actin staining (Alexa Fluor 488 Phalloidin, Life Technologies), performed on fixed cells, was executed according to the manufacturer’s procedure. .. PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA) as previously described [ ]. .. For transmigration assays, PBMCs were resuspended at 10x106 cells in 30 ml of Transendothelial migration buffer, TEM buffer (RPMI 1640 without phenol red + 1% bovine serum albumin + Hepes + L-glutamine + Na-pyruvate + MEM non-essential amino acids) and assayed within 2 h from phlebotomy.

    Article Title: Phase I Pharmacokinetic and Pharmacodynamic Evaluation of Combined Valproic Acid/Doxorubicin Treatment in Dogs with Spontaneous Cancer
    Article Snippet: Isolated peripheral blood mononuclear cells (PBMC) from pre- and post-VPA blood samples were stained for acetylated histones H3 using total histone H3 as a control. .. PBMC were isolated using Lymphocyte Separation Media [(LSM); Mediatech, Inc., Herndon, VA]. ..

    Article Title: FcRL4 Expression Identifies a Pro-inflammatory B Cell Subset in Viremic HIV-Infected Subjects
    Article Snippet: Antibodies Cells were stained with the following antibodies: FcRL4-APC (Biolegend), IL-6-PE, CD23-PE-Cy7, CD19-PE-Texas Red, CD10-Pe-Cy5, CD21-V450, CD27-AF700, phospho-p38-PE, phospho-Erk-AF647, phospho-p65-PE, phospho-C-Jun-FITC, (BD Biosciences), Sheep anti-rabbit IgG-DyLight 488 (Biolegend), purified hematopoietic cell kinase (Hck), and phospho-Hck (Abcam). .. Isolation, Purification, and TLR Stimulation of PBMCs PBMCs were isolated from whole blood using Ficoll (Lymphocyte® Cell Separation Media, Mediatech) gradient centrifugation. ..

    Article Title: Structural design of disialoganglioside GD2 and CD3-bispecific antibodies to redirect T cells for tumor therapy
    Article Snippet: Total release was assessed by lysis with 10% SDS (Sigma, St Louis, Mo), and background release was measured in the absence of effector cells . .. PBMC was isolated from healthy donor blood by lymphocyte separation medium (Mediatech Inc) centrifugation. .. Human T cells were purified by Pan T cell isolation kit (Miltenyi Biotec).

    Article Title: Omega-3 fatty acid supplementation decreases matrix metalloproteinase-9 production in relapsing-remitting multiple sclerosis
    Article Snippet: Blood samples for PBMC isolation were collected by venipuncture into herparinized Vacutainer® tubes. .. PBMC were isolated within 4 hours of blood draw (Lymphocyte separation media; Mediatech) and were incubated in PBS at 2 × 106 cells/ml at 37°C for 48hrs. ..

    Article Title: Type III IFNs are produced by and stimulate human plasmacytoid dendritic cells 1
    Article Snippet: In this study, mAb from clonal hybridoma cell line was used and its specificity was confirmed in flow cytometry with the use of CHO-derived cells overexpressing modified human IFN-λ (IL-28/29) receptor complex ( ) vs. non-transfected CHO cells (shown in ). .. PBMC were isolated by Ficoll-Hypaque (Mediatech, Inc, Herndon, VA) density centrifugation from fresh, heparinized blood obtained from healthy adult volunteers. ..

    Article Title: IL-6 blockade suppresses the blood-brain barrier disorder, leading to prevention of onset of NMOSD
    Article Snippet: .. PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA, USA) as previously described , and used in transmigration assays within 2 hours of phlebotomy. .. For transmigration assays, PBMCs were resuspended at 10 × 106 cells in 30 mL TEM buffer (RPMI 1640 without phenol red + 1% bovine serum albumin + 25 mM HEPES) and stained with Calcein AM (Thermo Fisher Scientific) prior to perfusion into the 3D flow chamber device following the manufacturer’s protocol (see below).

    Centrifugation:

    Article Title: Sphingosine 1 Phosphate at the Blood Brain Barrier: Can the Modulation of S1P Receptor 1 Influence the Response of Endothelial Cells and Astrocytes to Inflammatory Stimuli?
    Article Snippet: F-actin staining (Alexa Fluor 488 Phalloidin, Life Technologies), performed on fixed cells, was executed according to the manufacturer’s procedure. .. PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA) as previously described [ ]. .. For transmigration assays, PBMCs were resuspended at 10x106 cells in 30 ml of Transendothelial migration buffer, TEM buffer (RPMI 1640 without phenol red + 1% bovine serum albumin + Hepes + L-glutamine + Na-pyruvate + MEM non-essential amino acids) and assayed within 2 h from phlebotomy.

    Article Title: Structural design of disialoganglioside GD2 and CD3-bispecific antibodies to redirect T cells for tumor therapy
    Article Snippet: Total release was assessed by lysis with 10% SDS (Sigma, St Louis, Mo), and background release was measured in the absence of effector cells . .. PBMC was isolated from healthy donor blood by lymphocyte separation medium (Mediatech Inc) centrifugation. .. Human T cells were purified by Pan T cell isolation kit (Miltenyi Biotec).

    Article Title: Type III IFNs are produced by and stimulate human plasmacytoid dendritic cells 1
    Article Snippet: In this study, mAb from clonal hybridoma cell line was used and its specificity was confirmed in flow cytometry with the use of CHO-derived cells overexpressing modified human IFN-λ (IL-28/29) receptor complex ( ) vs. non-transfected CHO cells (shown in ). .. PBMC were isolated by Ficoll-Hypaque (Mediatech, Inc, Herndon, VA) density centrifugation from fresh, heparinized blood obtained from healthy adult volunteers. ..

    Article Title: IL-6 blockade suppresses the blood-brain barrier disorder, leading to prevention of onset of NMOSD
    Article Snippet: .. PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA, USA) as previously described , and used in transmigration assays within 2 hours of phlebotomy. .. For transmigration assays, PBMCs were resuspended at 10 × 106 cells in 30 mL TEM buffer (RPMI 1640 without phenol red + 1% bovine serum albumin + 25 mM HEPES) and stained with Calcein AM (Thermo Fisher Scientific) prior to perfusion into the 3D flow chamber device following the manufacturer’s protocol (see below).

    Gradient Centrifugation:

    Article Title: The Hepatitis B Vaccine Protects Re-Exposed Healthcare Workers, but Does Not Provide Sterilizing Immunity
    Article Snippet: .. PBMC were separated from heparin-anticoagulated blood by Ficoll-Histopaque (Mediatech, Manassas, VA) density gradient centrifugation, washed three times with phosphate buffered saline (PBS; Mediatech) as described , and cryopreserved in liquid nitrogen until use. .. Two hundred eighty-two 15-mer peptides were synthesized according to the HBV Galibert sequence (Genbank accession number ) with a 10 amino acid overlap (Mimotopes, Clayton, Australia) and resuspended in PBS containing 5% DMSO to generate one HBVcore (HBc) pool (41 peptides), two HBs pools (38 peptides each) and four HBVpolymerase (HBVpol) pools (three HBVpol pools consisting of 41 peptides each and one HBVpol pool consisting of 42 peptides).

    Article Title: FcRL4 Expression Identifies a Pro-inflammatory B Cell Subset in Viremic HIV-Infected Subjects
    Article Snippet: Antibodies Cells were stained with the following antibodies: FcRL4-APC (Biolegend), IL-6-PE, CD23-PE-Cy7, CD19-PE-Texas Red, CD10-Pe-Cy5, CD21-V450, CD27-AF700, phospho-p38-PE, phospho-Erk-AF647, phospho-p65-PE, phospho-C-Jun-FITC, (BD Biosciences), Sheep anti-rabbit IgG-DyLight 488 (Biolegend), purified hematopoietic cell kinase (Hck), and phospho-Hck (Abcam). .. Isolation, Purification, and TLR Stimulation of PBMCs PBMCs were isolated from whole blood using Ficoll (Lymphocyte® Cell Separation Media, Mediatech) gradient centrifugation. ..

    Purification:

    Article Title: FcRL4 Expression Identifies a Pro-inflammatory B Cell Subset in Viremic HIV-Infected Subjects
    Article Snippet: Antibodies Cells were stained with the following antibodies: FcRL4-APC (Biolegend), IL-6-PE, CD23-PE-Cy7, CD19-PE-Texas Red, CD10-Pe-Cy5, CD21-V450, CD27-AF700, phospho-p38-PE, phospho-Erk-AF647, phospho-p65-PE, phospho-C-Jun-FITC, (BD Biosciences), Sheep anti-rabbit IgG-DyLight 488 (Biolegend), purified hematopoietic cell kinase (Hck), and phospho-Hck (Abcam). .. Isolation, Purification, and TLR Stimulation of PBMCs PBMCs were isolated from whole blood using Ficoll (Lymphocyte® Cell Separation Media, Mediatech) gradient centrifugation. ..

    Incubation:

    Article Title: Omega-3 fatty acid supplementation decreases matrix metalloproteinase-9 production in relapsing-remitting multiple sclerosis
    Article Snippet: Blood samples for PBMC isolation were collected by venipuncture into herparinized Vacutainer® tubes. .. PBMC were isolated within 4 hours of blood draw (Lymphocyte separation media; Mediatech) and were incubated in PBS at 2 × 106 cells/ml at 37°C for 48hrs. ..

    Transmigration Assay:

    Article Title: IL-6 blockade suppresses the blood-brain barrier disorder, leading to prevention of onset of NMOSD
    Article Snippet: .. PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA, USA) as previously described , and used in transmigration assays within 2 hours of phlebotomy. .. For transmigration assays, PBMCs were resuspended at 10 × 106 cells in 30 mL TEM buffer (RPMI 1640 without phenol red + 1% bovine serum albumin + 25 mM HEPES) and stained with Calcein AM (Thermo Fisher Scientific) prior to perfusion into the 3D flow chamber device following the manufacturer’s protocol (see below).

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    Mediatech pbmc isolation peripheral blood mononuclear cells pbmcs
    S1P modulation reduces transmigration of <t>PBMCs</t> across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in <t>PBMC</t> migration after 1 hour compared to T I. *p
    Pbmc Isolation Peripheral Blood Mononuclear Cells Pbmcs, supplied by Mediatech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc isolation peripheral blood mononuclear cells pbmcs/product/Mediatech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmc isolation peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    86
    Mediatech peripheral blood mononuclear cells pbmcs
    S1P modulation reduces transmigration of <t>PBMCs</t> across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in <t>PBMC</t> migration after 1 hour compared to T I. *p
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Mediatech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Mediatech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    Image Search Results


    S1P modulation reduces transmigration of PBMCs across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in PBMC migration after 1 hour compared to T I. *p

    Journal: PLoS ONE

    Article Title: Sphingosine 1 Phosphate at the Blood Brain Barrier: Can the Modulation of S1P Receptor 1 Influence the Response of Endothelial Cells and Astrocytes to Inflammatory Stimuli?

    doi: 10.1371/journal.pone.0133392

    Figure Lengend Snippet: S1P modulation reduces transmigration of PBMCs across an in vitro BBB model under shear stress. When hBMVEC/hAST co-culture was exposed to T I in the presence of S1P (50 nM), pFTY (50 nM), or AUY (50 nM), the ability of PBMCs to migrate through the barrier is reduced if compared to the condition in which the barrier is exposed only to cytokines. Data are mean of 2 independent experiments and are expressed as percent reduction in PBMC migration after 1 hour compared to T I. *p

    Article Snippet: PBMC isolation Peripheral blood mononuclear cells (PBMCs) were isolated from fresh heparinized blood of healthy subjects by density centrifugation with Lymphocyte Separation Medium (Mediatech, Herndon, VA) as previously described [ ].

    Techniques: Transmigration Assay, In Vitro, Co-Culture Assay, Migration