Structured Review

Millipore blood mononuclear cells pbmcs
FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in <t>AIH</t> patients. <t>PBMCs</t> were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).
Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "The Imbalance between Foxp3+Tregs and Th1/Th17/Th22 Cells in Patients with Newly Diagnosed Autoimmune Hepatitis"

Article Title: The Imbalance between Foxp3+Tregs and Th1/Th17/Th22 Cells in Patients with Newly Diagnosed Autoimmune Hepatitis

Journal: Journal of Immunology Research

doi: 10.1155/2018/3753081

FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).
Figure Legend Snippet: FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Techniques Used: FACS, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Flow Cytometry, Cytometry

FACS analysis of the numbers of different subsets of circulating CD3 + CD4 + T cells and ELISA analysis of serum IL-10 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with FITC-anti-CD3, PE-Cy7-anti-CD25, and PerCP-anti-CD4 or isotype controls, fixed, and permeabilized, followed by intracellular staining with PE-anti-Foxp3. The frequency of CD3 + CD4 + CD25 − Foxp3 + and CD3 + CD4 + CD25 + Foxp3 + T cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + Foxp3 + T cells were calculated, according to the total numbers of PBMCs and the frequency of different types of CD3 + CD4 + Foxp3 + T cells. The concentrations of serum IL-10 in individual subjects were determined by ELISA. (a) Flow cytometry analysis; (b) the numbers of CD3 + CD4 + CD25 + Foxp3 + T cells; (c) the numbers of CD3 + CD 4+ CD25 − Foxp3 + T cells; (d) serum levels of IL-10. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood and the mean levels of serum IL-10 in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).
Figure Legend Snippet: FACS analysis of the numbers of different subsets of circulating CD3 + CD4 + T cells and ELISA analysis of serum IL-10 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with FITC-anti-CD3, PE-Cy7-anti-CD25, and PerCP-anti-CD4 or isotype controls, fixed, and permeabilized, followed by intracellular staining with PE-anti-Foxp3. The frequency of CD3 + CD4 + CD25 − Foxp3 + and CD3 + CD4 + CD25 + Foxp3 + T cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + Foxp3 + T cells were calculated, according to the total numbers of PBMCs and the frequency of different types of CD3 + CD4 + Foxp3 + T cells. The concentrations of serum IL-10 in individual subjects were determined by ELISA. (a) Flow cytometry analysis; (b) the numbers of CD3 + CD4 + CD25 + Foxp3 + T cells; (c) the numbers of CD3 + CD 4+ CD25 − Foxp3 + T cells; (d) serum levels of IL-10. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood and the mean levels of serum IL-10 in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Techniques Used: FACS, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Flow Cytometry, Cytometry

2) Product Images from "Replication and Meta-Analysis of GWAS Identified Susceptibility Loci in Kawasaki Disease Confirm the Importance of B Lymphoid Tyrosine Kinase (BLK) in Disease Susceptibility"

Article Title: Replication and Meta-Analysis of GWAS Identified Susceptibility Loci in Kawasaki Disease Confirm the Importance of B Lymphoid Tyrosine Kinase (BLK) in Disease Susceptibility

Journal: PLoS ONE

doi: 10.1371/journal.pone.0072037

B cell population in peripheral blood mononuclear cells (PBMC) and BLK expression in peripheral blood leukocytes (PBLs) induced at the acute stage of KD. (A) PBMCs were stained with anti-CD19 or anti-CD3 monoclonal antibodies, and the percentage of CD19 + B cells and CD3 + T cells in samples taken from a patient at acute and convalescent stages of KD and from a healthy control were determined by multicolor flow cytometry. (B) Levels of BLK expression were determined by real-time RT-PCR, and levels in KD patients were compared to those in fever controls. Values are expressed as mean ± standard error (SE). RNA was harvested from PBLs from KD patients at different stages of disease development or from age-matched fever controls. KD1, n = 20, before IVIG treatment (within 24 h before IVIG treatment); KD2, n = 12, after IVIG treatment (3–7 days after IVIG treatment); and KD3, n = 10, convalescence stage (3 weeks after IVIG treatment). FC, n = 19, fever controls.
Figure Legend Snippet: B cell population in peripheral blood mononuclear cells (PBMC) and BLK expression in peripheral blood leukocytes (PBLs) induced at the acute stage of KD. (A) PBMCs were stained with anti-CD19 or anti-CD3 monoclonal antibodies, and the percentage of CD19 + B cells and CD3 + T cells in samples taken from a patient at acute and convalescent stages of KD and from a healthy control were determined by multicolor flow cytometry. (B) Levels of BLK expression were determined by real-time RT-PCR, and levels in KD patients were compared to those in fever controls. Values are expressed as mean ± standard error (SE). RNA was harvested from PBLs from KD patients at different stages of disease development or from age-matched fever controls. KD1, n = 20, before IVIG treatment (within 24 h before IVIG treatment); KD2, n = 12, after IVIG treatment (3–7 days after IVIG treatment); and KD3, n = 10, convalescence stage (3 weeks after IVIG treatment). FC, n = 19, fever controls.

Techniques Used: Expressing, Staining, Flow Cytometry, Cytometry, Quantitative RT-PCR

3) Product Images from "Long-Lasting Enfuvirtide Carrier Pentasaccharide Conjugates with Potent Anti-Human Immunodeficiency Virus Type 1 Activity ▿"

Article Title: Long-Lasting Enfuvirtide Carrier Pentasaccharide Conjugates with Potent Anti-Human Immunodeficiency Virus Type 1 Activity ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00827-09

Antiviral activities (EC 50 s) of enfuvirtide-CP conjugates EP40111 and EP40112 against HIV-1 IIIB/LAI infecting PBMCs in the absence or the presence of 2.5 μM antithrombin. These results represent the means and standard deviations of three independent
Figure Legend Snippet: Antiviral activities (EC 50 s) of enfuvirtide-CP conjugates EP40111 and EP40112 against HIV-1 IIIB/LAI infecting PBMCs in the absence or the presence of 2.5 μM antithrombin. These results represent the means and standard deviations of three independent

Techniques Used:

4) Product Images from "Hepatocellular carcinoma cell sensitivity to Vγ9Vδ2 T lymphocyte-mediated killing is increased by zoledronate"

Article Title: Hepatocellular carcinoma cell sensitivity to Vγ9Vδ2 T lymphocyte-mediated killing is increased by zoledronate

Journal: International Journal of Oncology

doi: 10.3892/ijo.2016.3403

γδ T cell effects on Zol-treated HCC cell lines. (A) Maturation stage of γδ T cells, indicated by CD27 and CD45RA expression. Upper, representative plot of CD3 + TCRVγ9 + γδ T cells gated from healthy donor PBMCs. Lower, maturation of CD3 + TCRVγ9 + γδ T cells in 14-day cultures stimulated with PHA in the presence of IL-2 and IL-15. (B) Cytotoxic activity of γδ T cells towards target cells preincubated for 16 h in the presence or absence of 5 μM Zol. γδ T effector (E) cells were co-cultured for 4 h with indicated target (T) cells (1.0×10 4 cells/well) at the indicated E:T ratios. EJ1 cells, T2 cells, and PHA blast served as references. Data shown are representative of three independent experiments. (C) Cytokine production by γδ T cells (1.0×10 5 cells/well) after 24-h co-culture with the indicated target cells (5.0×10 4 cells/well). Data represent mean ± SD of triplicate cultures. (D) Proliferative response of γδ T cells (5.0×10 4 cells/well) co-cultured with irradiated (90 Gy) HCC cell lines (5.0×10 3 cells/well) for 72 h, as determined by [ 3 H]-thymidine incorporation assay. Data represent mean ± SD of triplicate cultures. Significance was determined by a one-way ANOVA with the Bonferroni post-hoc test; * P
Figure Legend Snippet: γδ T cell effects on Zol-treated HCC cell lines. (A) Maturation stage of γδ T cells, indicated by CD27 and CD45RA expression. Upper, representative plot of CD3 + TCRVγ9 + γδ T cells gated from healthy donor PBMCs. Lower, maturation of CD3 + TCRVγ9 + γδ T cells in 14-day cultures stimulated with PHA in the presence of IL-2 and IL-15. (B) Cytotoxic activity of γδ T cells towards target cells preincubated for 16 h in the presence or absence of 5 μM Zol. γδ T effector (E) cells were co-cultured for 4 h with indicated target (T) cells (1.0×10 4 cells/well) at the indicated E:T ratios. EJ1 cells, T2 cells, and PHA blast served as references. Data shown are representative of three independent experiments. (C) Cytokine production by γδ T cells (1.0×10 5 cells/well) after 24-h co-culture with the indicated target cells (5.0×10 4 cells/well). Data represent mean ± SD of triplicate cultures. (D) Proliferative response of γδ T cells (5.0×10 4 cells/well) co-cultured with irradiated (90 Gy) HCC cell lines (5.0×10 3 cells/well) for 72 h, as determined by [ 3 H]-thymidine incorporation assay. Data represent mean ± SD of triplicate cultures. Significance was determined by a one-way ANOVA with the Bonferroni post-hoc test; * P

Techniques Used: Expressing, Activity Assay, Cell Culture, Co-Culture Assay, Irradiation, Thymidine Incorporation Assay

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Article Title: Role of Bak in UV-induced apoptosis in skin cancer and abrogation by HPV E6 proteins
Article Snippet: The monoclonal antibodies used were Bak (Ab-2, Calbiochem), p53 (D01, ICRF), p21 (Santa Cruz), Ki67 (Dako), α-Tubulin (Calbiochem), and anti-HA (12CA5, ICRF).

Incubation:

Article Title: SPARCL1 suppresses metastasis in prostate cancer
Article Snippet: .. Membranes were incubated with chicken polyclonal antibody specific for human SPARCL1 (Abcam, Cambridge, United Kingdom) at a 1:2000 dilution, and then the membranes were re‐probed using mouse monoclonal antibody specific for human anti‐β‐actin (Sigma Chemical Company, Saint Louis, MO, USA). .. The human gene SPARCL1 ORF (RC207583, OriGene Technologies, Inc, Rockville, MD) was subcloned into pBMN‐I‐GFP (Addgene Inc., Cambridge, MA, USA) to obtain a pBMN‐SPARCL1‐I‐GFP plasmid.

Article Title: Conditional Site-Specific Integration into Human Chromosome 19 by Using a Ligand-Dependent Chimeric Adeno-Associated Virus/Rep Protein
Article Snippet: The polyclonal antiserum against Rep proteins was obtained by immunizing rabbits with purified recombinant Rep68 produced in Escherichia coli ( ) and recognizes all four species of Rep. To check the expression of the Hook gene product (sFv/PDGFR fusion protein) ( ) in the stable transfectants, total cellular extracts were prepared as described above, fractionated on an SDS–12% polyacrylamide gel, and transferred to nitrocellulose membranes. .. To detect the sFv/PDGFR fusion protein, the membranes were incubated first with monoclonal antibody 9E10.2 (dilution, 1:500), which recognizes the Myc.1 epitope tag ( ) present as a tandem repeat near the transmembrane domain of sFv/PDGFR , and then with an alkaline phosphatase-conjugated goat polyclonal anti-mouse IgG antiserum (Sigma no. A7434; dilution, 1:2,000). .. In all immunoblotting experiments, 5% nonfat dry milk in TBST (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 0.05% Tween 20) was used as a blocking agent and for diluting the various antibodies.

Inhibition:

Article Title: The Roles of Integrins in Function of Human Neutrophils after Their Migration through Endothelium into Interstitial Matrix
Article Snippet: The particles have a low fluorescence intensity in standard media or when adhered to the surface of neutrophils, but increase their fluorescence when taken into acidic phagocytic vacuoles. .. Inhibition of integrin function The following antibodies were used at 10μg/ml to block integrin function: rat IgG2a anti-human CD29/β1-integrin (Mab13; BD Pharmingen); mouse IgG2a anti-human CD18/β2-integrin (IB4; Calbiochem); mouse IgG1 anti-human CD61/β3-integrin (SZ21; Beckman Coulter); matched control antibodies, rat IgG2a, mouse IgG2a (both eBioscience), mouse IgG1 (DAKO). .. In some experiments, RGDS peptide (Arg-Gly-Asp-Ser, 0.5 mM; Sigma) or CT7010 (20 μM) was used instead of antibody.

Blocking Assay:

Article Title: The Roles of Integrins in Function of Human Neutrophils after Their Migration through Endothelium into Interstitial Matrix
Article Snippet: The particles have a low fluorescence intensity in standard media or when adhered to the surface of neutrophils, but increase their fluorescence when taken into acidic phagocytic vacuoles. .. Inhibition of integrin function The following antibodies were used at 10μg/ml to block integrin function: rat IgG2a anti-human CD29/β1-integrin (Mab13; BD Pharmingen); mouse IgG2a anti-human CD18/β2-integrin (IB4; Calbiochem); mouse IgG1 anti-human CD61/β3-integrin (SZ21; Beckman Coulter); matched control antibodies, rat IgG2a, mouse IgG2a (both eBioscience), mouse IgG1 (DAKO). .. In some experiments, RGDS peptide (Arg-Gly-Asp-Ser, 0.5 mM; Sigma) or CT7010 (20 μM) was used instead of antibody.

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  • 98
    Millipore pbmcs proliferation
    <t>MSCs</t> from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.
    Pbmcs Proliferation, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs proliferation/product/Millipore
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmcs proliferation - by Bioz Stars, 2021-03
    98/100 stars
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    99
    Millipore peripheral blood mononuclear cells
    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and <t>peripheral</t> <t>mononuclear</t> <t>blood</t> cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.
    Peripheral Blood Mononuclear Cells, supplied by Millipore, 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/peripheral blood mononuclear cells/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells - by Bioz Stars, 2021-03
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    Image Search Results


    MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Journal: Stem Cells Translational Medicine

    Article Title: A Proinflammatory Secretome Mediates the Impaired Immunopotency of Human Mesenchymal Stromal Cells in Elderly Patients with Atherosclerosis

    doi: 10.1002/sctm.16-0221

    Figure Lengend Snippet: MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Article Snippet: To assess the effect of MSCs on suppressing monocyte‐depleted PBMCs proliferation, PBMCs were labeled with 10 uM carboxyfluorescein succinimidyl ester (CFSE) (Sigma), stimulated with anti‐CD3/CD28 beads (1 bead per cell) (Dynabeads Human T‐Activator CD3/CD28, Life Technologies) and cultured for 4 days with MSCs.

    Techniques: Flow Cytometry, Cytometry, Derivative Assay

    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques:

    PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Isolation, Standard Deviation, Derivative Assay

    Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Journal: Cell Proliferation

    Article Title: Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor, et al. Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor

    doi: 10.1111/cpr.12858

    Figure Lengend Snippet: Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Article Snippet: Peripheral blood mononuclear cells were isolated from the whole blood of healthy donors by Ficoll density gradient centrifugation.

    Techniques: Isolation, Labeling, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Flow Cytometry

    Human and murine ILC2 gating strategy. A. Human ILC2s were isolated from peripheral blood of healthy donors PBMCs or umbilical cord blood samples CBMCs and stained with antibodies against CD45 and lineage markers. Human ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - CRTH2 + CD127 + cells. B. Murine ILC2s were isolated from mouse lungs treated with recombinant IL-33 protein (250ng/mouse, i.t. ) and stained with antibodies against CD45 and lineage markers as described in the Materials and Methods. Murine ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - T1/ST2 + cells. The purity of sorted ILC2s was determined to be greater than 95%.

    Journal: bioRxiv

    Article Title: Non-canonical Activation of Human Group 2 Innate Lymphoid Cells by TLR4 Signaling

    doi: 10.1101/2020.10.29.361345

    Figure Lengend Snippet: Human and murine ILC2 gating strategy. A. Human ILC2s were isolated from peripheral blood of healthy donors PBMCs or umbilical cord blood samples CBMCs and stained with antibodies against CD45 and lineage markers. Human ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - CRTH2 + CD127 + cells. B. Murine ILC2s were isolated from mouse lungs treated with recombinant IL-33 protein (250ng/mouse, i.t. ) and stained with antibodies against CD45 and lineage markers as described in the Materials and Methods. Murine ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - T1/ST2 + cells. The purity of sorted ILC2s was determined to be greater than 95%.

    Article Snippet: Peripheral or Cord Blood Mononuclear Cells (PBMCs or CBMCs) were isolated from diluted umbilical cord blood (1:2) by density gradient centrifugation using density gradient medium, Histopaque® (Sigma Aldrich) and SepMateTM 50 mL tubes (STEMCELL Technologies) ( ).

    Techniques: Isolation, Staining, Recombinant