human peripheral blood mononuclear cells pbmcs  (ATCC)


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  • 99
    Name:
    Primary Peripheral Blood Mononuclear Cells PBMC Normal Human
    Description:

    Catalog Number:
    pcs-800-011
    Price:
    None
    Applications:
    Applications for use include the study of immunology, infection, cancer, hematology, and t-cell suppression assay.
    Host:
    Homo sapiens, human
    Cell Type:
    Mononuclear
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    Structured Review

    ATCC human peripheral blood mononuclear cells pbmcs
    Gene expression of tumor-suppressor genes. RNA was isolated from peripheral blood mononuclear cells <t>(PBMCs)</t> and <t>K562</t> cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for tumor-suppressor genes. GAPDH was used a reference gene and data were normalized to PBMCs. *p

    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmcs/product/ATCC
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming"

    Article Title: Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming

    Journal: Turkish Journal of Hematology

    doi: 10.4274/tjh.2018.0106

    Gene expression of tumor-suppressor genes. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for tumor-suppressor genes. GAPDH was used a reference gene and data were normalized to PBMCs. *p
    Figure Legend Snippet: Gene expression of tumor-suppressor genes. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for tumor-suppressor genes. GAPDH was used a reference gene and data were normalized to PBMCs. *p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction

    Gene expression of reprogramming factors and pluripotency markers. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for A) reprogramming factors and B) pluripotency markers. GAPDH was used as a reference gene and data were normalized to PBMCs. *p
    Figure Legend Snippet: Gene expression of reprogramming factors and pluripotency markers. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for A) reprogramming factors and B) pluripotency markers. GAPDH was used as a reference gene and data were normalized to PBMCs. *p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction

    2) Product Images from "Analogs of the Frog-skin Antimicrobial Peptide Temporin 1Tb Exhibit a Wider Spectrum of Activity and a Stronger Antibiofilm Potential as Compared to the Parental Peptide"

    Article Title: Analogs of the Frog-skin Antimicrobial Peptide Temporin 1Tb Exhibit a Wider Spectrum of Activity and a Stronger Antibiofilm Potential as Compared to the Parental Peptide

    Journal: Frontiers in Chemistry

    doi: 10.3389/fchem.2017.00024

    Cytotoxicity of TB, TB_L1FK, and TB_KKG6A on human PBMCs (A) and A549 cells (B) after 24 h of incubation at 37°C, 5% CO 2 . The cytotoxic activity was evaluated by the PI flow cytometric assay. Cells incubated with culture medium only (100% cell viability) and cells treated with cycloheximide (0% cell viability) were used as controls. Data are reported as mean ± standard error of three independent experiments.
    Figure Legend Snippet: Cytotoxicity of TB, TB_L1FK, and TB_KKG6A on human PBMCs (A) and A549 cells (B) after 24 h of incubation at 37°C, 5% CO 2 . The cytotoxic activity was evaluated by the PI flow cytometric assay. Cells incubated with culture medium only (100% cell viability) and cells treated with cycloheximide (0% cell viability) were used as controls. Data are reported as mean ± standard error of three independent experiments.

    Techniques Used: Incubation, Activity Assay, Flow Cytometry

    3) Product Images from "Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming"

    Article Title: Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming

    Journal: Turkish Journal of Hematology

    doi: 10.4274/tjh.2018.0106

    Gene expression of tumor-suppressor genes. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for tumor-suppressor genes. GAPDH was used a reference gene and data were normalized to PBMCs. *p
    Figure Legend Snippet: Gene expression of tumor-suppressor genes. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for tumor-suppressor genes. GAPDH was used a reference gene and data were normalized to PBMCs. *p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction

    Gene expression of reprogramming factors and pluripotency markers. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for A) reprogramming factors and B) pluripotency markers. GAPDH was used as a reference gene and data were normalized to PBMCs. *p
    Figure Legend Snippet: Gene expression of reprogramming factors and pluripotency markers. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and K562 cells and quantitative real-time polymerase chain reaction was performed. Relative gene expression was plotted for A) reprogramming factors and B) pluripotency markers. GAPDH was used as a reference gene and data were normalized to PBMCs. *p

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction

    4) Product Images from "Endotoxin Conditioning Induces VCP/p97-mediated and Inducible Nitric-oxide Synthase-dependent Tyr284 Nitration in Protein Phosphatase 2A *"

    Article Title: Endotoxin Conditioning Induces VCP/p97-mediated and Inducible Nitric-oxide Synthase-dependent Tyr284 Nitration in Protein Phosphatase 2A *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.099788

    PP2Ac associates with VCP/p97. A , PP2Ac was immunoprecipitated ( IP ) from fractions 40–45 of DEAE-chromatography, and VCP/p97 was identified by liquid chromatography/MS. The black bars indicate positions of 23 peptides in the sequence that provided 35% coverage. B , reciprocal co-immunoprecipitation of the endogenous VCP/p97 and PP2Ac from RAW264.7 cells. Normal IgG was used as control ( Cont. ). Representative images from two independent experiments are shown. C , human PBMCs were conditioned with LPS (10 ng/ml) for 16 h. The levels of PP2Ac and VCP/p97 in whole cell ( WC ) extracts were analyzed by immunoblotting ( left panel ). VCP/p97 was immunoprecipitated, and PP2Ac and PP2A A subunit association was analyzed by immunoblotting ( right panel ). Representative images from three independent experiments are shown. D , Superose 12 gel chromatography of fractions 40–45 from DEAE chromatography from LPS-conditioned RAW264.7 cells. The proteins in individual fractions were detected by immunoblotting with indicated antibodies. Representative images from two independent experiments are shown. E , binding of purified PP2A to immobilized GST-VCP/p97. Top panel , anti-GST immunoblot showing GST and GST-VCP/p97 fusion protein. Bottom two panels , bound PP2Ac and PP2A A subunits were detected by immunoblotting. Representative images from two independent experiments are shown.
    Figure Legend Snippet: PP2Ac associates with VCP/p97. A , PP2Ac was immunoprecipitated ( IP ) from fractions 40–45 of DEAE-chromatography, and VCP/p97 was identified by liquid chromatography/MS. The black bars indicate positions of 23 peptides in the sequence that provided 35% coverage. B , reciprocal co-immunoprecipitation of the endogenous VCP/p97 and PP2Ac from RAW264.7 cells. Normal IgG was used as control ( Cont. ). Representative images from two independent experiments are shown. C , human PBMCs were conditioned with LPS (10 ng/ml) for 16 h. The levels of PP2Ac and VCP/p97 in whole cell ( WC ) extracts were analyzed by immunoblotting ( left panel ). VCP/p97 was immunoprecipitated, and PP2Ac and PP2A A subunit association was analyzed by immunoblotting ( right panel ). Representative images from three independent experiments are shown. D , Superose 12 gel chromatography of fractions 40–45 from DEAE chromatography from LPS-conditioned RAW264.7 cells. The proteins in individual fractions were detected by immunoblotting with indicated antibodies. Representative images from two independent experiments are shown. E , binding of purified PP2A to immobilized GST-VCP/p97. Top panel , anti-GST immunoblot showing GST and GST-VCP/p97 fusion protein. Bottom two panels , bound PP2Ac and PP2A A subunits were detected by immunoblotting. Representative images from two independent experiments are shown.

    Techniques Used: Immunoprecipitation, Chromatography, Liquid Chromatography, Mass Spectrometry, Sequencing, Binding Assay, Purification

    Related Articles

    Trypan Blue Exclusion Assay:

    Article Title: Antibacterial activity and cytotoxicity of multi-walled carbon nanotubes decorated with silver nanoparticles
    Article Snippet: .. Cytotoxicity test The effects of Ag-MWCNTs on the viability of mouse liver hepatocytes (AML 12; Chungnam University, Daejeon Korea) and human peripheral blood mononuclear cells (PBMCs) (American Type Culture Collection [ATCC], Manassas, VA, USA) were evaluated using a trypan blue exclusion assay and a LIVE/DEAD® Viability/Cytotoxicity Kit. .. Cultured AML 12 cells and human PBMCs were plated in six-well plates (1×105 cells per well) in Dulbecco’s Modified Eagle’s Medium (DMEM) and Roswell Park Memorial Institute medium (RPMI), respectively, each supplemented with 10% (v/v) fetal bovine serum and 1% sterile antibiotic.

    Isolation:

    Article Title: A New Family of Small-Molecule CD4-Mimetic Compounds Contacts Highly Conserved Aspartic Acid 368 of HIV-1 gp120 and Mediates Antibody-Dependent Cellular Cytotoxicity
    Article Snippet: .. Primary human peripheral blood mononuclear cells (PBMCs) and CD4+ T cells were isolated, activated, and cultured as previously described ( , ). ..

    Quantitation Assay:

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines
    Article Snippet: .. Receptor abundance quantitation Cryopreserved PBMCs (ATCC, PCS-800-011, lot#81115172) were thawed to room temperature and slowly diluted with 9 mL pre-warmed RPMI-1640 medium (Gibco, 11875-093) supplemented with 10% fetal bovine serum (FBS, Seradigm, 1500-500, lot#322B15). ..

    Labeling:

    Article Title: Nkx2‐5 Is Expressed in Atherosclerotic Plaques and Attenuates Development of Atherosclerosis in Apolipoprotein E–Deficient Mice
    Article Snippet: .. Fresh adult human PBMCs (PCS‐800‐011; ATCC) were labeled with calcein acetomethoxy (AM) dye: PBMCs were pelleted at 240g for 10 minutes, resuspended in 1 mL of culture medium with 2.5 μmol/L of calcein AM from the kit, and incubated at 37°C (5% CO2 ) for 30 minutes. .. PBMCs were then washed 3 times with HAEC media and added to HAEC cells (150 000 labeled PBMCs per chamber).

    Incubation:

    Article Title: Nkx2‐5 Is Expressed in Atherosclerotic Plaques and Attenuates Development of Atherosclerosis in Apolipoprotein E–Deficient Mice
    Article Snippet: .. Fresh adult human PBMCs (PCS‐800‐011; ATCC) were labeled with calcein acetomethoxy (AM) dye: PBMCs were pelleted at 240g for 10 minutes, resuspended in 1 mL of culture medium with 2.5 μmol/L of calcein AM from the kit, and incubated at 37°C (5% CO2 ) for 30 minutes. .. PBMCs were then washed 3 times with HAEC media and added to HAEC cells (150 000 labeled PBMCs per chamber).

    other:

    Article Title: Potency of Combining Eucalyptus camaldulensis subsp. camaldulensis with Low-Dose Cisplatin in A549 Human Lung Adenocarcinomas and MCF-7 Breast Adenocarcinoma
    Article Snippet: Peripheral Blood Mononuclear Cells (PBMC)Human peripheral blood mononuclear cells (PBMCs) obtained from ATCC, Manassas, VA, USA, comprising lymphocytes (B-cells, T-cells, and NK-cells), monocytes, and dendritic cells, were frequently used for the evaluation of immune responses.

    Cell Culture:

    Article Title: DNA-PKcs controls calcineurin mediated IL-2 production in T lymphocytes
    Article Snippet: .. Cell culture Human peripheral blood mononuclear cells (PBMC,) and Jurkat cells were purchased from ATCC (PCS-800-011, Manassas, VA). ..

    Article Title: Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming
    Article Snippet: .. Cell Culture Human CML cell line K562 and human peripheral blood mononuclear cells (PBMCs) were obtained from ATCC and Lonza, respectively. .. Cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum, 50 U/mL penicillin, 50 µg/mL streptomycin, and 1% L‐glutamine at 37 °C in 5% CO2 .

    Article Title: A New Family of Small-Molecule CD4-Mimetic Compounds Contacts Highly Conserved Aspartic Acid 368 of HIV-1 gp120 and Mediates Antibody-Dependent Cellular Cytotoxicity
    Article Snippet: .. Primary human peripheral blood mononuclear cells (PBMCs) and CD4+ T cells were isolated, activated, and cultured as previously described ( , ). ..

    Gradient Centrifugation:

    Article Title: Critical Role for the NLRP3 Inflammasome in Mediating IL-1β Production in Shigella sonnei-Infected Macrophages
    Article Snippet: .. THP-1 macrophages were differentiated from THP-1 monocytes by treatment with 50 nM PMA for 48 h. Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood from healthy volunteers by density gradient centrifugation using Histopaque-1077 , and all experimental protocols were performed in accordance with the guidelines and regulations provided and accepted by the Institutional Review Board of the Tri-Service General Hospital, National Defense Medical Center and the volunteers' informed consent (TSGH-IRB-2-106-05-190 and TSGH-IRB-2-106-05-009). .. Mouse primary bone marrow derived macrophages (BMDM) were prepared from bone marrow collected from C57BL/6 mouse femur and tibia by differentiating in the M-CSF containing medium for 7 days.

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  • 99
    ATCC mutans atcc 25175 stimulated pbmcs
    Auto-aggregation of Lactobacillus brevis BBE-Y52 and coaggregation of BBE-Y52 with oral pathogens. Auto-aggregation (lines) of individual strain and coaggregation (dot lines) of BBE-Y52 with oral pathogens. Solid symbols: square, L . brevis BBE-Y52; circle, Porphyromonas gingivalis GIM1.851; upper triangle, Fusobacterium nucleatum CGMCC 1.2528; lower triangle, Streptococcus <t>mutans</t> ATCC 25175; Open symbols: right triangle, L . brevis BBE-Y52 + S . mutans ATCC 25175; diamond, L . brevis BBE-Y52 + P. gingivalis GIM1.851; star, L . brevis BBE-Y52 + F . nucleatum CGMCC 1.2528
    Mutans Atcc 25175 Stimulated Pbmcs, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mutans atcc 25175 stimulated pbmcs/product/ATCC
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mutans atcc 25175 stimulated pbmcs - by Bioz Stars, 2020-09
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    99
    ATCC peripheral blood mononuclear cells pbmc human peripheral blood mononuclear cells pbmcs
    Cell viability of peripheral blood mononuclear cells <t>(PBMC)</t> after treatment for 24 h. ( A ) Cell viability of <t>PBMC</t> cells after treatment with the ethanolic extract of E. camaldulensis for 24 h. ( B ) Cell viability of PBMC cells after treatment with increasing concentrations of CDDP for 24 h. ( C ) Cell viability of PBMC cells after treatment with the aqueous extract of E. camaldulensis for 24 h. ( D ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 75 µg/mL). ( E ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 150 µg/mL) (* p
    Peripheral Blood Mononuclear Cells Pbmc Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmc human peripheral blood mononuclear cells pbmcs/product/ATCC
    Average 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmc human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Auto-aggregation of Lactobacillus brevis BBE-Y52 and coaggregation of BBE-Y52 with oral pathogens. Auto-aggregation (lines) of individual strain and coaggregation (dot lines) of BBE-Y52 with oral pathogens. Solid symbols: square, L . brevis BBE-Y52; circle, Porphyromonas gingivalis GIM1.851; upper triangle, Fusobacterium nucleatum CGMCC 1.2528; lower triangle, Streptococcus mutans ATCC 25175; Open symbols: right triangle, L . brevis BBE-Y52 + S . mutans ATCC 25175; diamond, L . brevis BBE-Y52 + P. gingivalis GIM1.851; star, L . brevis BBE-Y52 + F . nucleatum CGMCC 1.2528

    Journal: BMC Microbiology

    Article Title: Characterization of a Lactobacillus brevis strain with potential oral probiotic properties

    doi: 10.1186/s12866-018-1369-3

    Figure Lengend Snippet: Auto-aggregation of Lactobacillus brevis BBE-Y52 and coaggregation of BBE-Y52 with oral pathogens. Auto-aggregation (lines) of individual strain and coaggregation (dot lines) of BBE-Y52 with oral pathogens. Solid symbols: square, L . brevis BBE-Y52; circle, Porphyromonas gingivalis GIM1.851; upper triangle, Fusobacterium nucleatum CGMCC 1.2528; lower triangle, Streptococcus mutans ATCC 25175; Open symbols: right triangle, L . brevis BBE-Y52 + S . mutans ATCC 25175; diamond, L . brevis BBE-Y52 + P. gingivalis GIM1.851; star, L . brevis BBE-Y52 + F . nucleatum CGMCC 1.2528

    Article Snippet: For the double-challenge, S . mutans ATCC 25175-stimulated PBMCs (2 × 106 cell/mL) were incubated either alone or with oral lactobacilli (108 CFU/mL) for 24 h prior to cytokine analysis [ ].

    Techniques:

    Antimicrobial activity of Lactobacillus brevis BBE-Y52 against Streptococcus mutans ATCC 25175. Control, growth of S . mutans ATCC 25175 with addition of lactic acid (pH 4.2) in the well; L . brevis BBE-Y52, inhibition of S . mutans ATCC 25175 by the supernatant of L . brevis BBE-Y52

    Journal: BMC Microbiology

    Article Title: Characterization of a Lactobacillus brevis strain with potential oral probiotic properties

    doi: 10.1186/s12866-018-1369-3

    Figure Lengend Snippet: Antimicrobial activity of Lactobacillus brevis BBE-Y52 against Streptococcus mutans ATCC 25175. Control, growth of S . mutans ATCC 25175 with addition of lactic acid (pH 4.2) in the well; L . brevis BBE-Y52, inhibition of S . mutans ATCC 25175 by the supernatant of L . brevis BBE-Y52

    Article Snippet: For the double-challenge, S . mutans ATCC 25175-stimulated PBMCs (2 × 106 cell/mL) were incubated either alone or with oral lactobacilli (108 CFU/mL) for 24 h prior to cytokine analysis [ ].

    Techniques: Activity Assay, Inhibition

    Biofilm formation by Lactobacillus brevis BBE-Y52 and S . mutans ATCC 25175. Black bars, white bars and grey bars represent the biofilm formed by individual Streptococcus mutans ATCC 25175, L . brevis BBE-Y52, and both strains, respectively. * and ** represent significant ( P

    Journal: BMC Microbiology

    Article Title: Characterization of a Lactobacillus brevis strain with potential oral probiotic properties

    doi: 10.1186/s12866-018-1369-3

    Figure Lengend Snippet: Biofilm formation by Lactobacillus brevis BBE-Y52 and S . mutans ATCC 25175. Black bars, white bars and grey bars represent the biofilm formed by individual Streptococcus mutans ATCC 25175, L . brevis BBE-Y52, and both strains, respectively. * and ** represent significant ( P

    Article Snippet: For the double-challenge, S . mutans ATCC 25175-stimulated PBMCs (2 × 106 cell/mL) were incubated either alone or with oral lactobacilli (108 CFU/mL) for 24 h prior to cytokine analysis [ ].

    Techniques:

    Cell viability of peripheral blood mononuclear cells (PBMC) after treatment for 24 h. ( A ) Cell viability of PBMC cells after treatment with the ethanolic extract of E. camaldulensis for 24 h. ( B ) Cell viability of PBMC cells after treatment with increasing concentrations of CDDP for 24 h. ( C ) Cell viability of PBMC cells after treatment with the aqueous extract of E. camaldulensis for 24 h. ( D ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 75 µg/mL). ( E ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 150 µg/mL) (* p

    Journal: Medicines

    Article Title: Potency of Combining Eucalyptus camaldulensis subsp. camaldulensis with Low-Dose Cisplatin in A549 Human Lung Adenocarcinomas and MCF-7 Breast Adenocarcinoma

    doi: 10.3390/medicines7080040

    Figure Lengend Snippet: Cell viability of peripheral blood mononuclear cells (PBMC) after treatment for 24 h. ( A ) Cell viability of PBMC cells after treatment with the ethanolic extract of E. camaldulensis for 24 h. ( B ) Cell viability of PBMC cells after treatment with increasing concentrations of CDDP for 24 h. ( C ) Cell viability of PBMC cells after treatment with the aqueous extract of E. camaldulensis for 24 h. ( D ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 75 µg/mL). ( E ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 150 µg/mL) (* p

    Article Snippet: Peripheral Blood Mononuclear Cells (PBMC)Human peripheral blood mononuclear cells (PBMCs) obtained from ATCC, Manassas, VA, USA, comprising lymphocytes (B-cells, T-cells, and NK-cells), monocytes, and dendritic cells, were frequently used for the evaluation of immune responses.

    Techniques:

    Stenotic aortic valve immune infiltrate is a smaller source of nucleotide-degrading ecto-nucleotidases but a larger of adenosine deaminase. Simplified protocol of stenotic aortic valve cell isolation, including endothelial cells (first step of isolation, vWF positive or CD31 high positive), interstitial cells (second step of isolation; Vimentin positive, Vim+) and immune cells (first and second step of isolation; CD45 positive, CD45+) ( a ). Flow cytometry analysis of CD45 positive cells (immune cells) as a percentage of total isolated cells after first step of isolation (cells located in the upper layers of the valve) and second step of isolation (cells located in the deeper layers of the valve) ( b ). The composition of stenotic aor tic valve immune infiltrate ( c ) expressed as a percentage (%) of total CD45+ cells, including T helper cells (CD45+, CD4+), T cytotoxic cells (CD45+, CD8+), B cells (CD45+, CD19+), monocytes/macrophages (CD45+, CD11b+, CD14+) and granulocytes (CD45+, CD11b int , CD14−). The rates of ATP hydrolysis, AMP hydrolysis and adenosine deamination on the surface of human monocyte/macrophages (SC; d ) and human peripheral blood mononuclear cells (PBMC = lymphocytes; e ) and in the presence of ecto-enzyme inhibitors. Results are shown as mean ± SEM; n = 5–9, * p

    Journal: Clinical Research in Cardiology

    Article Title: Nucleotide ecto-enzyme metabolic pattern and spatial distribution in calcific aortic valve disease; its relation to pathological changes and clinical presentation

    doi: 10.1007/s00392-019-01495-x

    Figure Lengend Snippet: Stenotic aortic valve immune infiltrate is a smaller source of nucleotide-degrading ecto-nucleotidases but a larger of adenosine deaminase. Simplified protocol of stenotic aortic valve cell isolation, including endothelial cells (first step of isolation, vWF positive or CD31 high positive), interstitial cells (second step of isolation; Vimentin positive, Vim+) and immune cells (first and second step of isolation; CD45 positive, CD45+) ( a ). Flow cytometry analysis of CD45 positive cells (immune cells) as a percentage of total isolated cells after first step of isolation (cells located in the upper layers of the valve) and second step of isolation (cells located in the deeper layers of the valve) ( b ). The composition of stenotic aor tic valve immune infiltrate ( c ) expressed as a percentage (%) of total CD45+ cells, including T helper cells (CD45+, CD4+), T cytotoxic cells (CD45+, CD8+), B cells (CD45+, CD19+), monocytes/macrophages (CD45+, CD11b+, CD14+) and granulocytes (CD45+, CD11b int , CD14−). The rates of ATP hydrolysis, AMP hydrolysis and adenosine deamination on the surface of human monocyte/macrophages (SC; d ) and human peripheral blood mononuclear cells (PBMC = lymphocytes; e ) and in the presence of ecto-enzyme inhibitors. Results are shown as mean ± SEM; n = 5–9, * p

    Article Snippet: Isolated human peripheral blood mononuclear cells and monocyte/macrophage cells (SC line, ATCC, cat. CRL-9855) that were used at passage 4, were plated in 24-well cell culture plate at a density 0.2 × 106 per well in a total volume of 1 mL HBSS.

    Techniques: Cell Isolation, Isolation, Flow Cytometry

    S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Journal: Frontiers in Immunology

    Article Title: Critical Role for the NLRP3 Inflammasome in Mediating IL-1β Production in Shigella sonnei-Infected Macrophages

    doi: 10.3389/fimmu.2020.01115

    Figure Lengend Snippet: S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Article Snippet: THP-1 macrophages were differentiated from THP-1 monocytes by treatment with 50 nM PMA for 48 h. Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood from healthy volunteers by density gradient centrifugation using Histopaque-1077 , and all experimental protocols were performed in accordance with the guidelines and regulations provided and accepted by the Institutional Review Board of the Tri-Service General Hospital, National Defense Medical Center and the volunteers' informed consent (TSGH-IRB-2-106-05-190 and TSGH-IRB-2-106-05-009).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot