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cd3 t cell isolation kit 896  (Miltenyi Biotec)


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    Miltenyi Biotec cd3 t cell isolation kit 896
    Cd3 T Cell Isolation Kit 896, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 544 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd3+t+cell/CD3+MicroBeads%2C+human/pm42374531-323-24-29
    Average 96 stars, based on 544 article reviews
    cd3 t cell isolation kit 896 - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Isolation:

    Article Title: A Comparative assessment of T-Cell response of Healthy donors and Acute Graft-versus-Host-Disease Patients: Customizing immune monitoring platform
    Article Snippet: Peripheral blood (PB) was collected from healthy individuals and patients in a sterile sodium heparin-coated vacutainer (BD Biosciences, US) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll density gradient centrifugation according to a standardized protocol [ ]. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with Phytohemagglutinin (PHA) (Sigma, USA) at concentrations ranging from 1.0μg/ml to 10μg/ml with IL-2 (50IU/ml-500IU/ml) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37?C, 5% CO 2 for 24, 48 and 72 hours.

    Article Title: Extracellular vesicles derived from nasopharyngeal carcinoma induce the emergence of mature regulatory dendritic cells using a galectin-9 dependent mechanism.
    Article Snippet: .. Human monocyte and T cell isolations Monocytes and total CD3 T cell were isolated from PBMCs using, respectively, a positive magnetic selection CD14+ and CD3+ isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions with an average purity of 95% and 75%, respectively. . Monocytes derived dendritic cells (moDC) generation Monocytes (106 cells/mL) were cultivated in complete medium (RPMI 1640, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) with 10% clarified FBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 50 μg/mL of gentamycin (Thermo Fisher Scientific,Waltham,MA,USA) in 6-well plates (Corning Incorporated,NY,USA). .. To allowmonocytes differentiation into immature DCs (iDC), GM-CSF (25 ng/mL) and IL4 (10 ng/mL) (PeproTech Inc, Rocky Hill, NJ, USA) were added to the medium.

    Article Title: Dose-response immunomodulatory effects of Mesenchymal stem cell-derived culture-conditioned media in acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA), and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer's instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park's Memorial Institute (RPMI)-1640 medium supplemented with l -glutamine (Thermo Fisher Scientific, USA), 10 % FBS (Thermo Fisher Scientific, USA), 1 % antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37֯C, 5 % CO 2 for 48 h [ ].

    Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease
    Article Snippet: The buffy coat containing mononuclear cells was collected and washed with 1X PBS (Thermo Fisher Scientific, USA). .. CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions. .. CD3 + T-cell was labeled with 1μM Cell Trace TM CFSE dye (BD Biosciences, USA) for 20 minutes at 37C and followed by activation of CD3 + T-cell with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI) medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS, 1% antibiotic-antimycotic solution at 37C, 5% CO 2 for 48 hours.

    Article Title: Dose-Response Immunomodulatory Effects of Mesenchymal Stem Cells-Derived Culture-Conditioned Media in Acute Graft-versus-Host-Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37°C, 5% CO 2 for 48 hours ( ).

    Article Title: Hypoxia-induced metabolic and apoptotic reprogramming enhances immunomodulation in Wharton’s jelly mesenchymal stem cells
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGVHD patients ( n = 25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. CD3 + T cell was isolated using the Pan T cell isolation kit (Miltenyi Biotec, USA) through negative selection and cultured for 48 h in the presence of PHA (1 μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA), following our previously established protocols described elsewhere. ..

    Article Title: Hypoxia-Mediated Molecular Interactions of Tissue-Specific Mesenchymal Stem Cells Drive Metabolic Reprogramming and Immunomodulation in Acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGvHD patients (n=25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. The isolation of peripheral blood mononuclear cells (PBMNCs) was performed through Ficoll density gradient centrifugation (María Del Pilar De la Rosa-Ruiz et al, 2019) and subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) for 48 hours according to our established protocols ( ). .. For co-culture experiments, tissue-specific (BM, WJ) MSCs (MSCs, MSCs HYP ) were treated with mitomycin-C at a concentration of 15μg/ml (Thermo Fisher Scientific, USA) for 1 hour in 1X LG-DMEM incomplete medium.

    Selection:

    Article Title: A Comparative assessment of T-Cell response of Healthy donors and Acute Graft-versus-Host-Disease Patients: Customizing immune monitoring platform
    Article Snippet: Peripheral blood (PB) was collected from healthy individuals and patients in a sterile sodium heparin-coated vacutainer (BD Biosciences, US) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll density gradient centrifugation according to a standardized protocol [ ]. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with Phytohemagglutinin (PHA) (Sigma, USA) at concentrations ranging from 1.0μg/ml to 10μg/ml with IL-2 (50IU/ml-500IU/ml) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37?C, 5% CO 2 for 24, 48 and 72 hours.

    Article Title: Extracellular vesicles derived from nasopharyngeal carcinoma induce the emergence of mature regulatory dendritic cells using a galectin-9 dependent mechanism.
    Article Snippet: .. Human monocyte and T cell isolations Monocytes and total CD3 T cell were isolated from PBMCs using, respectively, a positive magnetic selection CD14+ and CD3+ isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions with an average purity of 95% and 75%, respectively. . Monocytes derived dendritic cells (moDC) generation Monocytes (106 cells/mL) were cultivated in complete medium (RPMI 1640, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) with 10% clarified FBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 50 μg/mL of gentamycin (Thermo Fisher Scientific,Waltham,MA,USA) in 6-well plates (Corning Incorporated,NY,USA). .. To allowmonocytes differentiation into immature DCs (iDC), GM-CSF (25 ng/mL) and IL4 (10 ng/mL) (PeproTech Inc, Rocky Hill, NJ, USA) were added to the medium.

    Article Title: Dose-response immunomodulatory effects of Mesenchymal stem cell-derived culture-conditioned media in acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA), and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer's instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park's Memorial Institute (RPMI)-1640 medium supplemented with l -glutamine (Thermo Fisher Scientific, USA), 10 % FBS (Thermo Fisher Scientific, USA), 1 % antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37֯C, 5 % CO 2 for 48 h [ ].

    Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease
    Article Snippet: The buffy coat containing mononuclear cells was collected and washed with 1X PBS (Thermo Fisher Scientific, USA). .. CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions. .. CD3 + T-cell was labeled with 1μM Cell Trace TM CFSE dye (BD Biosciences, USA) for 20 minutes at 37C and followed by activation of CD3 + T-cell with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI) medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS, 1% antibiotic-antimycotic solution at 37C, 5% CO 2 for 48 hours.

    Article Title: Dual Preconditioned Mesenchymal Stem Cells-Derived Culture Conditioned Media Augment Immunomodulation and Drive Metabolic Reprogramming in Acute Graft-versus-Host Disease
    Article Snippet: .. Subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), following the manufacturer’s protocol. .. The isolated CD3 + T-cell were stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) using our standardized protocol ( ).

    Article Title: Dose-Response Immunomodulatory Effects of Mesenchymal Stem Cells-Derived Culture-Conditioned Media in Acute Graft-versus-Host-Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37°C, 5% CO 2 for 48 hours ( ).

    Article Title: Hypoxia-induced metabolic and apoptotic reprogramming enhances immunomodulation in Wharton’s jelly mesenchymal stem cells
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGVHD patients ( n = 25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. CD3 + T cell was isolated using the Pan T cell isolation kit (Miltenyi Biotec, USA) through negative selection and cultured for 48 h in the presence of PHA (1 μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA), following our previously established protocols described elsewhere. ..

    Article Title: Hypoxia-Mediated Molecular Interactions of Tissue-Specific Mesenchymal Stem Cells Drive Metabolic Reprogramming and Immunomodulation in Acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGvHD patients (n=25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. The isolation of peripheral blood mononuclear cells (PBMNCs) was performed through Ficoll density gradient centrifugation (María Del Pilar De la Rosa-Ruiz et al, 2019) and subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) for 48 hours according to our established protocols ( ). .. For co-culture experiments, tissue-specific (BM, WJ) MSCs (MSCs, MSCs HYP ) were treated with mitomycin-C at a concentration of 15μg/ml (Thermo Fisher Scientific, USA) for 1 hour in 1X LG-DMEM incomplete medium.

    Cell Isolation:

    Article Title: A Comparative assessment of T-Cell response of Healthy donors and Acute Graft-versus-Host-Disease Patients: Customizing immune monitoring platform
    Article Snippet: Peripheral blood (PB) was collected from healthy individuals and patients in a sterile sodium heparin-coated vacutainer (BD Biosciences, US) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll density gradient centrifugation according to a standardized protocol [ ]. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with Phytohemagglutinin (PHA) (Sigma, USA) at concentrations ranging from 1.0μg/ml to 10μg/ml with IL-2 (50IU/ml-500IU/ml) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37?C, 5% CO 2 for 24, 48 and 72 hours.

    Article Title: Dose-response immunomodulatory effects of Mesenchymal stem cell-derived culture-conditioned media in acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA), and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer's instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park's Memorial Institute (RPMI)-1640 medium supplemented with l -glutamine (Thermo Fisher Scientific, USA), 10 % FBS (Thermo Fisher Scientific, USA), 1 % antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37֯C, 5 % CO 2 for 48 h [ ].

    Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease
    Article Snippet: The buffy coat containing mononuclear cells was collected and washed with 1X PBS (Thermo Fisher Scientific, USA). .. CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions. .. CD3 + T-cell was labeled with 1μM Cell Trace TM CFSE dye (BD Biosciences, USA) for 20 minutes at 37C and followed by activation of CD3 + T-cell with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI) medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS, 1% antibiotic-antimycotic solution at 37C, 5% CO 2 for 48 hours.

    Article Title: Dual Preconditioned Mesenchymal Stem Cells-Derived Culture Conditioned Media Augment Immunomodulation and Drive Metabolic Reprogramming in Acute Graft-versus-Host Disease
    Article Snippet: .. Subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), following the manufacturer’s protocol. .. The isolated CD3 + T-cell were stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) using our standardized protocol ( ).

    Article Title: Dose-Response Immunomodulatory Effects of Mesenchymal Stem Cells-Derived Culture-Conditioned Media in Acute Graft-versus-Host-Disease
    Article Snippet: Peripheral blood (PB) was collected from aGVHD patients in a sterile sodium heparin-coated vacutainer (Becton Dickinson, USA) and the peripheral blood mononuclear cells (PBMNCs) were isolated using Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) density gradient centrifugation as described elsewhere. .. CD3 + T-cell was then isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA), following the manufacturer’s instructions. .. Subsequently, CD3 + T-cell was stimulated with 1.0 μg/ml of Phytohemagglutinin (PHA) (Sigma-Aldrich, Saint Louis, MO, USA) and 50 IU/ml of IL-2 (Thermo Fisher Scientific, USA) in 1X Rosewell Park’s Memorial Institute (RPMI)-1640 medium supplemented with L-glutamine (Thermo Fisher Scientific, USA), 10% FBS (Thermo Fisher Scientific, USA), 1% antibiotic-antimycotic solution (Thermo Fisher Scientific, USA) at 37°C, 5% CO 2 for 48 hours ( ).

    Article Title: Hypoxia-induced metabolic and apoptotic reprogramming enhances immunomodulation in Wharton’s jelly mesenchymal stem cells
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGVHD patients ( n = 25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. CD3 + T cell was isolated using the Pan T cell isolation kit (Miltenyi Biotec, USA) through negative selection and cultured for 48 h in the presence of PHA (1 μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA), following our previously established protocols described elsewhere. ..

    Article Title: Hypoxia-Mediated Molecular Interactions of Tissue-Specific Mesenchymal Stem Cells Drive Metabolic Reprogramming and Immunomodulation in Acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGvHD patients (n=25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. The isolation of peripheral blood mononuclear cells (PBMNCs) was performed through Ficoll density gradient centrifugation (María Del Pilar De la Rosa-Ruiz et al, 2019) and subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) for 48 hours according to our established protocols ( ). .. For co-culture experiments, tissue-specific (BM, WJ) MSCs (MSCs, MSCs HYP ) were treated with mitomycin-C at a concentration of 15μg/ml (Thermo Fisher Scientific, USA) for 1 hour in 1X LG-DMEM incomplete medium.

    Derivative Assay:

    Article Title: Extracellular vesicles derived from nasopharyngeal carcinoma induce the emergence of mature regulatory dendritic cells using a galectin-9 dependent mechanism.
    Article Snippet: .. Human monocyte and T cell isolations Monocytes and total CD3 T cell were isolated from PBMCs using, respectively, a positive magnetic selection CD14+ and CD3+ isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions with an average purity of 95% and 75%, respectively. . Monocytes derived dendritic cells (moDC) generation Monocytes (106 cells/mL) were cultivated in complete medium (RPMI 1640, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) with 10% clarified FBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 50 μg/mL of gentamycin (Thermo Fisher Scientific,Waltham,MA,USA) in 6-well plates (Corning Incorporated,NY,USA). .. To allowmonocytes differentiation into immature DCs (iDC), GM-CSF (25 ng/mL) and IL4 (10 ng/mL) (PeproTech Inc, Rocky Hill, NJ, USA) were added to the medium.

    Cell Culture:

    Article Title: Hypoxia-induced metabolic and apoptotic reprogramming enhances immunomodulation in Wharton’s jelly mesenchymal stem cells
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGVHD patients ( n = 25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. CD3 + T cell was isolated using the Pan T cell isolation kit (Miltenyi Biotec, USA) through negative selection and cultured for 48 h in the presence of PHA (1 μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA), following our previously established protocols described elsewhere. ..

    Gradient Centrifugation:

    Article Title: Hypoxia-Mediated Molecular Interactions of Tissue-Specific Mesenchymal Stem Cells Drive Metabolic Reprogramming and Immunomodulation in Acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGvHD patients (n=25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. The isolation of peripheral blood mononuclear cells (PBMNCs) was performed through Ficoll density gradient centrifugation (María Del Pilar De la Rosa-Ruiz et al, 2019) and subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) for 48 hours according to our established protocols ( ). .. For co-culture experiments, tissue-specific (BM, WJ) MSCs (MSCs, MSCs HYP ) were treated with mitomycin-C at a concentration of 15μg/ml (Thermo Fisher Scientific, USA) for 1 hour in 1X LG-DMEM incomplete medium.

    Staining:

    Article Title: Hypoxia-Mediated Molecular Interactions of Tissue-Specific Mesenchymal Stem Cells Drive Metabolic Reprogramming and Immunomodulation in Acute Graft-versus-Host Disease
    Article Snippet: Peripheral blood (PB) was collected from grade II-IV aGvHD patients (n=25) in sterile sodium heparin-coated vacutainers (BD Biosciences, US). .. The isolation of peripheral blood mononuclear cells (PBMNCs) was performed through Ficoll density gradient centrifugation (María Del Pilar De la Rosa-Ruiz et al, 2019) and subsequently, CD3 + T-cell were extracted from the PBMNCs through negative selection using a Pan T cell isolation kit (Miltenyi Biotec, USA), stained with 1 μM cell Trace TM carboxyfluorescein succinimidyl ester (CFSE) dye (BD Biosciences, USA) and activated with PHA (1μg/ml) (Sigma, USA) and IL-2 (50IU/ml) (Thermo Fisher Scientific, USA) for 48 hours according to our established protocols ( ). .. For co-culture experiments, tissue-specific (BM, WJ) MSCs (MSCs, MSCs HYP ) were treated with mitomycin-C at a concentration of 15μg/ml (Thermo Fisher Scientific, USA) for 1 hour in 1X LG-DMEM incomplete medium.



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    cd3 t cells  (Miltenyi Biotec)
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    Miltenyi Biotec cd3 t cells
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