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  • 99
    Thermo Fisher cfse
    Cfse, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 31001 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cfse/product/Thermo Fisher
    Average 99 stars, based on 31001 article reviews
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    cfse - by Bioz Stars, 2020-09
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    99
    Millipore cfse
    Expansion of antigen-specific T cells ex vivo and in vivo by G-DCs. a , b Antigen-specific CD8+ T-cell stimulation by G-DCs, live DCs, and glutaraldehyde-fixed DCs ex vivo. Error bars represent mean ± SEM. n = 3. c , d Antigen-specific CD8+ T-cell stimulation by G-DCs and live DCs in vivo. Error bars represent mean ± SEM. ( n = 4 to 8). e , f G-DCs and non-gelated DCs were stored in <t>PBS</t> at 4 °C for 21 days (Day 21 G-DC and Day 21 DCs). In vivo antigen-specific CD8+ T-cell stimulation was examined with day 21 G-DC, day 21 DCs, and day 21 G-DC structurally disrupted by ultrasonication (homogenized G-DCs). T-cell expansion was monitored on day 6. <t>CFSE-labeled</t> T-cell groups (gray) are shown as negative controls. Error bars represent mean ± SEM, n = 3
    Cfse, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cfse/product/Millipore
    Average 99 stars, based on 3187 article reviews
    Price from $9.99 to $1999.99
    cfse - by Bioz Stars, 2020-09
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    99
    Thermo Fisher celltrace cfse cell proliferation kit
    Polarization of STAT1 at the NK-target cell interface. (A) Murine wild-type <t>LAK</t> cells were co-incubated with <t>CFSE-labeled</t> Stat1 −/− v-abl + leukemic cells (green) in a ratio of 1:1 for 30 min prior to the immunofluorescent staining of STAT1 (red). Nuclear staining with DAPI (blue) was included as control. Scale bars: 10 µm. (B) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT1 (red) and F-actin (yellow). Upon tumor cell challenge STAT1 remained cytoplasmic and was partially found in the NK-immunological synapse, as it co-localized with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm. (C) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT5 (red) and F-actin (yellow). Upon tumor cell challenge STAT5 remained evenly distributed over the NK cell and was not recruited to the NK immunological synpase as it did not co-localize with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm.
    Celltrace Cfse Cell Proliferation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4793 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/celltrace cfse cell proliferation kit/product/Thermo Fisher
    Average 99 stars, based on 4793 article reviews
    Price from $9.99 to $1999.99
    celltrace cfse cell proliferation kit - by Bioz Stars, 2020-09
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    99
    Thermo Fisher carboxyfluorescein diacetate succinimidyl ester cfse
    The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. <t>Carboxyfluorescein</t> diacetate <t>succinimidyl</t> ester <t>(CFSE)-labelled</t> OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.
    Carboxyfluorescein Diacetate Succinimidyl Ester Cfse, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carboxyfluorescein diacetate succinimidyl ester cfse/product/Thermo Fisher
    Average 99 stars, based on 2168 article reviews
    Price from $9.99 to $1999.99
    carboxyfluorescein diacetate succinimidyl ester cfse - by Bioz Stars, 2020-09
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    99
    Thermo Fisher vybrant cfse cell tracer kit
    The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. <t>Carboxyfluorescein</t> diacetate <t>succinimidyl</t> ester <t>(CFSE)-labelled</t> OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.
    Vybrant Cfse Cell Tracer Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vybrant cfse cell tracer kit/product/Thermo Fisher
    Average 99 stars, based on 77 article reviews
    Price from $9.99 to $1999.99
    vybrant cfse cell tracer kit - by Bioz Stars, 2020-09
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    98
    BioLegend cfse cell division tracker kit
    The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. <t>Carboxyfluorescein</t> diacetate <t>succinimidyl</t> ester <t>(CFSE)-labelled</t> OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.
    Cfse Cell Division Tracker Kit, supplied by BioLegend, used in various techniques. Bioz Stars score: 98/100, based on 264 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cfse cell division tracker kit/product/BioLegend
    Average 98 stars, based on 264 article reviews
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    cfse cell division tracker kit - by Bioz Stars, 2020-09
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    96
    Millipore carboxyfluorescein diacetate succinimidyl ester cfse
    The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. <t>Carboxyfluorescein</t> diacetate <t>succinimidyl</t> ester <t>(CFSE)-labelled</t> OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.
    Carboxyfluorescein Diacetate Succinimidyl Ester Cfse, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carboxyfluorescein diacetate succinimidyl ester cfse/product/Millipore
    Average 96 stars, based on 153 article reviews
    Price from $9.99 to $1999.99
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    93
    Becton Dickinson cfse
    Volume is an implicit microenvironment factor driving MSC potency. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. Proliferation was measured through flow <t>cytometry</t> and <t>CFSE</t> staining; bar graphs represent mean +/− SD of 3 samples. (A ) Bar graphs of normalized proliferation versus culture volume conditions. ( B ) Bar graphs of normalized cytokine secretion versus culture volume conditions. Two-group significance comparisons were performed with a student’s T-test; n.s., no significance; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.
    Cfse, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 2835 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cfse/product/Becton Dickinson
    Average 93 stars, based on 2835 article reviews
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    cfse - by Bioz Stars, 2020-09
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    93
    Dojindo Labs cfse
    Knockdown of Erbin promotes proliferation and metastasis of cervical cancer cells in nude mice. ( a , b ) In all, 5 × 10 6 <t>HeLa/NC</t> and HeLa/Erbin-sh cells were subcutaneously injected into the right flanks of six-to-seven-week old female Balb/C athymic nude mice. The mice were monitored daily, and tumor volumes estimated according to the formula volume=length × width 2 /2. At the end of the experiments, the mice were killed. The primary tumors were dissected and weighted. ( c , d ) HeLa/NC and HeLa/Erbin-sh cells were labeled with <t>CFSE</t> and then implanted into the peritoneal cavity of nude mice. At 72 h after implantation, the apoptotic rates of the tumor cells from peritoneal cavity were determined by FACS analysis of CFSE and Annexin V-phycoerythrin double-positive cells. ( e ) In all, 5 × 10 6 HeLa/NC and HeLa/Erbin-sh cells were implanted into the peritoneal cavity of nude mice. After implantation for 56 days, the peritoneal cavities were dissected and photographed. ( f ) The expression of Erbin in human cervical cancer tissues was analyzed by immunohistochemistry with the antibody against Erbin using a tissue microarray containing cervical cancer and normal cervical tissues.
    Cfse, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 93/100, based on 215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cfse/product/Dojindo Labs
    Average 93 stars, based on 215 article reviews
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    cfse - by Bioz Stars, 2020-09
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    Image Search Results


    Expansion of antigen-specific T cells ex vivo and in vivo by G-DCs. a , b Antigen-specific CD8+ T-cell stimulation by G-DCs, live DCs, and glutaraldehyde-fixed DCs ex vivo. Error bars represent mean ± SEM. n = 3. c , d Antigen-specific CD8+ T-cell stimulation by G-DCs and live DCs in vivo. Error bars represent mean ± SEM. ( n = 4 to 8). e , f G-DCs and non-gelated DCs were stored in PBS at 4 °C for 21 days (Day 21 G-DC and Day 21 DCs). In vivo antigen-specific CD8+ T-cell stimulation was examined with day 21 G-DC, day 21 DCs, and day 21 G-DC structurally disrupted by ultrasonication (homogenized G-DCs). T-cell expansion was monitored on day 6. CFSE-labeled T-cell groups (gray) are shown as negative controls. Error bars represent mean ± SEM, n = 3

    Journal: Nature Communications

    Article Title: Intracellular hydrogelation preserves fluid and functional cell membrane interfaces for biological interactions

    doi: 10.1038/s41467-019-09049-5

    Figure Lengend Snippet: Expansion of antigen-specific T cells ex vivo and in vivo by G-DCs. a , b Antigen-specific CD8+ T-cell stimulation by G-DCs, live DCs, and glutaraldehyde-fixed DCs ex vivo. Error bars represent mean ± SEM. n = 3. c , d Antigen-specific CD8+ T-cell stimulation by G-DCs and live DCs in vivo. Error bars represent mean ± SEM. ( n = 4 to 8). e , f G-DCs and non-gelated DCs were stored in PBS at 4 °C for 21 days (Day 21 G-DC and Day 21 DCs). In vivo antigen-specific CD8+ T-cell stimulation was examined with day 21 G-DC, day 21 DCs, and day 21 G-DC structurally disrupted by ultrasonication (homogenized G-DCs). T-cell expansion was monitored on day 6. CFSE-labeled T-cell groups (gray) are shown as negative controls. Error bars represent mean ± SEM, n = 3

    Article Snippet: OT-I cells were stained with carboxyfluorescein diacetate succinimidyl ester (CFSE) by incubating the cells with PBS containing 5 μM of CFSE (Sigma-Aldrich, #21888) at 37 °C for 5 min.

    Techniques: Ex Vivo, In Vivo, Cell Stimulation, Labeling

    Polarization of STAT1 at the NK-target cell interface. (A) Murine wild-type LAK cells were co-incubated with CFSE-labeled Stat1 −/− v-abl + leukemic cells (green) in a ratio of 1:1 for 30 min prior to the immunofluorescent staining of STAT1 (red). Nuclear staining with DAPI (blue) was included as control. Scale bars: 10 µm. (B) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT1 (red) and F-actin (yellow). Upon tumor cell challenge STAT1 remained cytoplasmic and was partially found in the NK-immunological synapse, as it co-localized with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm. (C) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT5 (red) and F-actin (yellow). Upon tumor cell challenge STAT5 remained evenly distributed over the NK cell and was not recruited to the NK immunological synpase as it did not co-localize with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm.

    Journal: Oncoimmunology

    Article Title: Novel non-canonical role of STAT1 in Natural Killer cell cytotoxicity

    doi: 10.1080/2162402X.2016.1186314

    Figure Lengend Snippet: Polarization of STAT1 at the NK-target cell interface. (A) Murine wild-type LAK cells were co-incubated with CFSE-labeled Stat1 −/− v-abl + leukemic cells (green) in a ratio of 1:1 for 30 min prior to the immunofluorescent staining of STAT1 (red). Nuclear staining with DAPI (blue) was included as control. Scale bars: 10 µm. (B) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT1 (red) and F-actin (yellow). Upon tumor cell challenge STAT1 remained cytoplasmic and was partially found in the NK-immunological synapse, as it co-localized with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm. (C) Murine wild-type LAK cells were co-incubated with CFSE-labeled human Jurkat cells (green) for 30 min prior to the immunofluorescent staining of STAT5 (red) and F-actin (yellow). Upon tumor cell challenge STAT5 remained evenly distributed over the NK cell and was not recruited to the NK immunological synpase as it did not co-localize with F-actin. Nuclear staining with DAPI (blue) was included as control. Scale bars: 5 µm.

    Article Snippet: 10 wild-type LAK cells were co-incubated in a ratio of 1:1 with CFSE-labeled (2.5 µM; CellTrace CFSE Cell Proliferation Kit, Molecular Probes) murine Stat1 −/− v-abl+ leukemic or human Jurkat cells in RPMI complete medium.

    Techniques: Incubation, Labeling, Staining

    Lama5 regulates CD4 + T cell and Treg entry into LNs via αDG and α6 integrin. ( A ) CFSE-labeled iTregs (2 × 10 6 ) and 2 × 10 6 eFluor 670–labeled CD4 + cells transferred i.v. to Lama5-KO or WT mice. After 16 hours, LNs were stained for ER-TR7 and with DAPI and analyzed for the transferred cells. Left panels: Representative whole-section images (original magnification, ×20); arrowheads indicate HEVs. Scale bars: 200 μm (left) and 50 μm (right). Right panels: Quantification of naive CD4 + T cells and iTregs in the CR and HEVs ( n = 30). ( B and C ) T cells pretreated with anti-αDG (2.5 μg mAb/10 6 cells, isotype IgM) or anti–α6 integrin (itg) (5 μg mAb/10 6 cells, isotype IgG) before transfer. After 16 hours, LNs were harvested for immunohistochemistry and flow cytometry. ( B ) Upper panels: Gating strategy. Lower panels: Number of transferred naive CD4 + T cells (eFluor 670 + ) and iTregs (CFSE + ) per 1 × 10 6 total CD4 + T cells or total Foxp3 + cells ( n = 6). ( C ) Upper panels: Representative scanning images (original magnification, ×20). LNs were stained for ER-TR7 and with DAPI and analyzed for transferred cells. Scale bar: 50 μm. Lower panels: Quantification of transferred naive CD4 + T cells (eFluor 670 + ) and iTregs (CFSE + ) in the CR and HEVs ( n = 30). Data (mean ± SEM) are from 3 independent experiments with 3 mice/group. For immunohistochemistry, 5 LNs/mouse, 3 sections/LN, and 3–5 fields/section. * P

    Journal: The Journal of Clinical Investigation

    Article Title: The lymph node stromal laminin α5 shapes alloimmunity

    doi: 10.1172/JCI135099

    Figure Lengend Snippet: Lama5 regulates CD4 + T cell and Treg entry into LNs via αDG and α6 integrin. ( A ) CFSE-labeled iTregs (2 × 10 6 ) and 2 × 10 6 eFluor 670–labeled CD4 + cells transferred i.v. to Lama5-KO or WT mice. After 16 hours, LNs were stained for ER-TR7 and with DAPI and analyzed for the transferred cells. Left panels: Representative whole-section images (original magnification, ×20); arrowheads indicate HEVs. Scale bars: 200 μm (left) and 50 μm (right). Right panels: Quantification of naive CD4 + T cells and iTregs in the CR and HEVs ( n = 30). ( B and C ) T cells pretreated with anti-αDG (2.5 μg mAb/10 6 cells, isotype IgM) or anti–α6 integrin (itg) (5 μg mAb/10 6 cells, isotype IgG) before transfer. After 16 hours, LNs were harvested for immunohistochemistry and flow cytometry. ( B ) Upper panels: Gating strategy. Lower panels: Number of transferred naive CD4 + T cells (eFluor 670 + ) and iTregs (CFSE + ) per 1 × 10 6 total CD4 + T cells or total Foxp3 + cells ( n = 6). ( C ) Upper panels: Representative scanning images (original magnification, ×20). LNs were stained for ER-TR7 and with DAPI and analyzed for transferred cells. Scale bar: 50 μm. Lower panels: Quantification of transferred naive CD4 + T cells (eFluor 670 + ) and iTregs (CFSE + ) in the CR and HEVs ( n = 30). Data (mean ± SEM) are from 3 independent experiments with 3 mice/group. For immunohistochemistry, 5 LNs/mouse, 3 sections/LN, and 3–5 fields/section. * P

    Article Snippet: CD4+ T cells were stained with a Cell Trace CFSE Cell Proliferation Kit (Thermo Fisher Scientific).

    Techniques: Labeling, Mouse Assay, Staining, Immunohistochemistry, Flow Cytometry

    The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. Carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.

    Journal: Innate Immunity

    Article Title: The second and third amino acids of Pam2 lipopeptides are key for the proliferation of cytotoxic T cells

    doi: 10.1177/1753425918777598

    Figure Lengend Snippet: The Aa following Pam2Cys determines the cross-priming activity in bone marrow–derived DC (BMDC). (a) and (b) BMDCs were stimulated with Pam2LPs in the presence of OVA. Carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled OT-I cells were co-cultured with the BMDCs. After 60 h, the degree of OT-I proliferation was assessed by flow cytometry. (a) OT-I cells were gated as CD8α + TCR Vβ5.1,5.2 + population. The histograms of representative samples are shown. (b) The result including all samples is shown. The results are representative of two independent experiments.

    Article Snippet: Carboxyfluorescein diacetate succinimidyl ester (CFSE) was purchased from Molecular Probes (C1157).

    Techniques: Activity Assay, Derivative Assay, Cell Culture, Flow Cytometry, Cytometry

    Down-regulation of NKG2D by the MICA-129Met and the MICA-129Val isoforms on CD8 + T cells and impairment of subsequent co-stimulation via NKG2D NKG2D expression on purified CD8 + T cells exposed to L-MICA-129Met ( n = 19) or L-MICA-129Val clones ( n = 19) for 0, 4, and 24 h was analyzed by flow cytometry. CD8 + T cells (2.5 × 10 5 ) were co-cultured with 5 × 10 4 target cells and analyzed for NKG2D expression after gating on CD3 + CD8 + T cells. The means and SD of the MFI of NKG2D (left panel) and of the percentage of NKG2D + CD8 + T cells (right panel) are displayed. Differences between the groups were analyzed by repeated measures ANOVA, and the P -values are indicated. Purified CD8 + T cells were cultured on plate-bound anti-NKG2D (1 μg/ml) or isotype control (mIgG 1 ) for 24 h before the NKG2D expression was measured by flow cytometry. Means and SD of MFI (upper panel) and percentage of NKG2D + cells (lower panel) are shown ( n = 6). Differences between the groups were analyzed by t -tests, and the P -values are indicated. These CD8 + T cells were subsequently CFSE-stained and cultured on plates coated with anti-CD3 (0.005 μg/ml) in combination with anti-CD28 (0.5 μg/ml) as a positive control or anti-NKG2D (0.5 μg/ml). Proliferation was measured after 60 h by flow cytometry. Untreated CFSE-stained CD8 + T cells are included for comparison. The percentage of proliferating cells and the MFI for CFSE are indicated. The MFI of CFSE in unstimulated CD8 + T cells (control) was set to 100% in individual experiments ( n = 6), and the relative decrease due to proliferation was calculated. Means + SD are shown. Significant differences (Wilcoxon test) between CD8 + T cells pre-exposed to anti-NKG2D and isotype control (mIgG 1 ) were found in these experiments at anti-CD3 concentrations of 0.01 and 0.005 μg/ml.

    Journal: EMBO Molecular Medicine

    Article Title: The MICA-129 dimorphism affects NKG2D signaling and outcome of hematopoietic stem cell transplantation

    doi: 10.15252/emmm.201505246

    Figure Lengend Snippet: Down-regulation of NKG2D by the MICA-129Met and the MICA-129Val isoforms on CD8 + T cells and impairment of subsequent co-stimulation via NKG2D NKG2D expression on purified CD8 + T cells exposed to L-MICA-129Met ( n = 19) or L-MICA-129Val clones ( n = 19) for 0, 4, and 24 h was analyzed by flow cytometry. CD8 + T cells (2.5 × 10 5 ) were co-cultured with 5 × 10 4 target cells and analyzed for NKG2D expression after gating on CD3 + CD8 + T cells. The means and SD of the MFI of NKG2D (left panel) and of the percentage of NKG2D + CD8 + T cells (right panel) are displayed. Differences between the groups were analyzed by repeated measures ANOVA, and the P -values are indicated. Purified CD8 + T cells were cultured on plate-bound anti-NKG2D (1 μg/ml) or isotype control (mIgG 1 ) for 24 h before the NKG2D expression was measured by flow cytometry. Means and SD of MFI (upper panel) and percentage of NKG2D + cells (lower panel) are shown ( n = 6). Differences between the groups were analyzed by t -tests, and the P -values are indicated. These CD8 + T cells were subsequently CFSE-stained and cultured on plates coated with anti-CD3 (0.005 μg/ml) in combination with anti-CD28 (0.5 μg/ml) as a positive control or anti-NKG2D (0.5 μg/ml). Proliferation was measured after 60 h by flow cytometry. Untreated CFSE-stained CD8 + T cells are included for comparison. The percentage of proliferating cells and the MFI for CFSE are indicated. The MFI of CFSE in unstimulated CD8 + T cells (control) was set to 100% in individual experiments ( n = 6), and the relative decrease due to proliferation was calculated. Means + SD are shown. Significant differences (Wilcoxon test) between CD8 + T cells pre-exposed to anti-NKG2D and isotype control (mIgG 1 ) were found in these experiments at anti-CD3 concentrations of 0.01 and 0.005 μg/ml.

    Article Snippet: In other experiments, purified CD8+ T cells were labeled by the dye CFSE (C-1157, Invitrogen, Karlsruhe, Germany) before culture on the coated plates.

    Techniques: Expressing, Purification, Clone Assay, Flow Cytometry, Cytometry, Cell Culture, Staining, Positive Control

    Volume is an implicit microenvironment factor driving MSC potency. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. Proliferation was measured through flow cytometry and CFSE staining; bar graphs represent mean +/− SD of 3 samples. (A ) Bar graphs of normalized proliferation versus culture volume conditions. ( B ) Bar graphs of normalized cytokine secretion versus culture volume conditions. Two-group significance comparisons were performed with a student’s T-test; n.s., no significance; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.

    Journal: Scientific Reports

    Article Title: Therapeutic Delivery Specifications Identified Through Compartmental Analysis of a Mesenchymal Stromal Cell-Immune Reaction

    doi: 10.1038/s41598-018-24971-2

    Figure Lengend Snippet: Volume is an implicit microenvironment factor driving MSC potency. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. Proliferation was measured through flow cytometry and CFSE staining; bar graphs represent mean +/− SD of 3 samples. (A ) Bar graphs of normalized proliferation versus culture volume conditions. ( B ) Bar graphs of normalized cytokine secretion versus culture volume conditions. Two-group significance comparisons were performed with a student’s T-test; n.s., no significance; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.

    Article Snippet: Flow cytometry was used to assess CFSE dilution associated with proliferation (LSRII, BD Biosciences, Franklin Lakes, NJ).

    Techniques: Flow Cytometry, Cytometry, Staining

    PBMC culture duration with MSCs is critical to overall immunosuppressive effect. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. MSC transwell inserts were removed after 1, 2, and 3 days co-culture initiation to time duration. Proliferation was measured through flow cytometry and CFSE staining; bar graphs and data points represent mean +/− SD of 3 samples. ( A ) Bar graphs of normalized proliferation versus time exposure to MSCs. ( B ) Bar graphs of normalized cytokine secretion versus time exposure to MSCs. Investigating cytokine profiling, we see concordant associations seen in previous sections. Longer exposure to MSCs results in greater suppression of inflammatory cytokines and vice versa for short exposures. Correlative lines were generated using a linear regression.

    Journal: Scientific Reports

    Article Title: Therapeutic Delivery Specifications Identified Through Compartmental Analysis of a Mesenchymal Stromal Cell-Immune Reaction

    doi: 10.1038/s41598-018-24971-2

    Figure Lengend Snippet: PBMC culture duration with MSCs is critical to overall immunosuppressive effect. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. MSC transwell inserts were removed after 1, 2, and 3 days co-culture initiation to time duration. Proliferation was measured through flow cytometry and CFSE staining; bar graphs and data points represent mean +/− SD of 3 samples. ( A ) Bar graphs of normalized proliferation versus time exposure to MSCs. ( B ) Bar graphs of normalized cytokine secretion versus time exposure to MSCs. Investigating cytokine profiling, we see concordant associations seen in previous sections. Longer exposure to MSCs results in greater suppression of inflammatory cytokines and vice versa for short exposures. Correlative lines were generated using a linear regression.

    Article Snippet: Flow cytometry was used to assess CFSE dilution associated with proliferation (LSRII, BD Biosciences, Franklin Lakes, NJ).

    Techniques: Co-Culture Assay, Flow Cytometry, Cytometry, Staining, Generated

    MSC:PBMC cross communication is critical as shown by the use of a protein transport inhibitor. PBMCs were treated for 24 hours with Brefeldin A prior to the start of co-culture; +BA indicates with brefeldin A pretreatment, -BA indicates without brefeldin A pretreatment. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. Proliferation was measured through flow cytometry and CFSE staining; bar graphs represent mean +/− SD of 3 samples. ( A ) Bar graphs of normalized proliferation versus Brefeldin A conditions. ( B ) Bar graphs of normalized cytokine secretion versus Brefeldin A conditions. Two-group significance comparisons were performed with a student’s T-test; n.s., no significance; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.

    Journal: Scientific Reports

    Article Title: Therapeutic Delivery Specifications Identified Through Compartmental Analysis of a Mesenchymal Stromal Cell-Immune Reaction

    doi: 10.1038/s41598-018-24971-2

    Figure Lengend Snippet: MSC:PBMC cross communication is critical as shown by the use of a protein transport inhibitor. PBMCs were treated for 24 hours with Brefeldin A prior to the start of co-culture; +BA indicates with brefeldin A pretreatment, -BA indicates without brefeldin A pretreatment. PBMC proliferation was attained through stimulation with ConA and IL2 for a period of 4 days. Proliferation was measured through flow cytometry and CFSE staining; bar graphs represent mean +/− SD of 3 samples. ( A ) Bar graphs of normalized proliferation versus Brefeldin A conditions. ( B ) Bar graphs of normalized cytokine secretion versus Brefeldin A conditions. Two-group significance comparisons were performed with a student’s T-test; n.s., no significance; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.

    Article Snippet: Flow cytometry was used to assess CFSE dilution associated with proliferation (LSRII, BD Biosciences, Franklin Lakes, NJ).

    Techniques: Co-Culture Assay, Flow Cytometry, Cytometry, Staining

    Pharmacological assessment of MSC immunosuppression with perturbation and regression analysis. PBMC proliferation was measured using flow cytometry and CFSE staining after stimulation with ConA and IL-2 for a period of 4 days. ( A ) Density plot of CFSE dilution; clear definition between proliferative generations (up to 5) is apparent. ( B ) Dose response curve of MSC suppression of T cell activation; data points represent mean +/− SD of 3 samples. Six ratios of MSCs were co-cultured with 1.5 M PBMCs to generate a full dose response curve (1:5, 1:10, 1:50, 1:100, 1:500, and 1:1000). This curve is fit by a pharmacologic dose response regression (Equation 2 ) with strong fit (R 2 = 0.99). ( C ) Independent variable assessment was performed by doubling the number of PBMCs; data points represent mean +/− SD of 3 samples. ( top ) Two distinct curves form using the metric of ratio showing poor universal applicably of this metric with a nearly 3-fold difference between IC50 values. ( middle bottom ) Implementing a cells/well and cell/mL approach, we find greatly improved agreement between these curves with a IC50 value differences less than 1.5-fold. ( D ) Each independent variable was then assessed again several matched studies to determine broad applicability using regression derived from 1C (Equation 2 ; regression values found in Table S1 ); each data point represent a distinct value from literature. Pharmacologic curves were derived from a non-linear, 4 parameter regression; correlative lines were generated using a linear regression.

    Journal: Scientific Reports

    Article Title: Therapeutic Delivery Specifications Identified Through Compartmental Analysis of a Mesenchymal Stromal Cell-Immune Reaction

    doi: 10.1038/s41598-018-24971-2

    Figure Lengend Snippet: Pharmacological assessment of MSC immunosuppression with perturbation and regression analysis. PBMC proliferation was measured using flow cytometry and CFSE staining after stimulation with ConA and IL-2 for a period of 4 days. ( A ) Density plot of CFSE dilution; clear definition between proliferative generations (up to 5) is apparent. ( B ) Dose response curve of MSC suppression of T cell activation; data points represent mean +/− SD of 3 samples. Six ratios of MSCs were co-cultured with 1.5 M PBMCs to generate a full dose response curve (1:5, 1:10, 1:50, 1:100, 1:500, and 1:1000). This curve is fit by a pharmacologic dose response regression (Equation 2 ) with strong fit (R 2 = 0.99). ( C ) Independent variable assessment was performed by doubling the number of PBMCs; data points represent mean +/− SD of 3 samples. ( top ) Two distinct curves form using the metric of ratio showing poor universal applicably of this metric with a nearly 3-fold difference between IC50 values. ( middle bottom ) Implementing a cells/well and cell/mL approach, we find greatly improved agreement between these curves with a IC50 value differences less than 1.5-fold. ( D ) Each independent variable was then assessed again several matched studies to determine broad applicability using regression derived from 1C (Equation 2 ; regression values found in Table S1 ); each data point represent a distinct value from literature. Pharmacologic curves were derived from a non-linear, 4 parameter regression; correlative lines were generated using a linear regression.

    Article Snippet: Flow cytometry was used to assess CFSE dilution associated with proliferation (LSRII, BD Biosciences, Franklin Lakes, NJ).

    Techniques: Flow Cytometry, Cytometry, Staining, Activation Assay, Cell Culture, Derivative Assay, Generated

    Knockdown of Erbin promotes proliferation and metastasis of cervical cancer cells in nude mice. ( a , b ) In all, 5 × 10 6 HeLa/NC and HeLa/Erbin-sh cells were subcutaneously injected into the right flanks of six-to-seven-week old female Balb/C athymic nude mice. The mice were monitored daily, and tumor volumes estimated according to the formula volume=length × width 2 /2. At the end of the experiments, the mice were killed. The primary tumors were dissected and weighted. ( c , d ) HeLa/NC and HeLa/Erbin-sh cells were labeled with CFSE and then implanted into the peritoneal cavity of nude mice. At 72 h after implantation, the apoptotic rates of the tumor cells from peritoneal cavity were determined by FACS analysis of CFSE and Annexin V-phycoerythrin double-positive cells. ( e ) In all, 5 × 10 6 HeLa/NC and HeLa/Erbin-sh cells were implanted into the peritoneal cavity of nude mice. After implantation for 56 days, the peritoneal cavities were dissected and photographed. ( f ) The expression of Erbin in human cervical cancer tissues was analyzed by immunohistochemistry with the antibody against Erbin using a tissue microarray containing cervical cancer and normal cervical tissues.

    Journal: Oncogenesis

    Article Title: Deficiency of Erbin induces resistance of cervical cancer cells to anoikis in a STAT3-dependent manner

    doi: 10.1038/oncsis.2013.18

    Figure Lengend Snippet: Knockdown of Erbin promotes proliferation and metastasis of cervical cancer cells in nude mice. ( a , b ) In all, 5 × 10 6 HeLa/NC and HeLa/Erbin-sh cells were subcutaneously injected into the right flanks of six-to-seven-week old female Balb/C athymic nude mice. The mice were monitored daily, and tumor volumes estimated according to the formula volume=length × width 2 /2. At the end of the experiments, the mice were killed. The primary tumors were dissected and weighted. ( c , d ) HeLa/NC and HeLa/Erbin-sh cells were labeled with CFSE and then implanted into the peritoneal cavity of nude mice. At 72 h after implantation, the apoptotic rates of the tumor cells from peritoneal cavity were determined by FACS analysis of CFSE and Annexin V-phycoerythrin double-positive cells. ( e ) In all, 5 × 10 6 HeLa/NC and HeLa/Erbin-sh cells were implanted into the peritoneal cavity of nude mice. After implantation for 56 days, the peritoneal cavities were dissected and photographed. ( f ) The expression of Erbin in human cervical cancer tissues was analyzed by immunohistochemistry with the antibody against Erbin using a tissue microarray containing cervical cancer and normal cervical tissues.

    Article Snippet: In vivo anoikis assays HeLa cells stably expressing control shRNA (HeLa/NC) or Erbin shRNA (HeLa/Erbin-sh) were labeled by CFSE (Dojindo Laboratories, Dojindo, Kumamoto, Japan) according to the manufacturer's instruction.

    Techniques: Mouse Assay, Injection, Labeling, FACS, Expressing, Immunohistochemistry, Microarray