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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and <t>PEVio770-CD45RA</t> staining.
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Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and PEVio770-CD45RA staining.

Journal: Cell Transplantation

Article Title: A novel GMP-manufactured medicinal product candidate composed of NK and γδ T cells as adjunct immunotherapy for hematopoietic stem cell transplantation

doi: 10.1177/09636897251374248

Figure Lengend Snippet: Flow cytometric analysis of cell populations during the process. Panel A. Number of cells and proportion of populations obtained at the different steps of the process starting from buffy coat (BC) or leukapheresis (LA). Panels B–D. One representative experiment using LA (#3) as staring material is shown. Panel B. CD14 + monocytes (a), CD19 + B cells (b) CD3 + T cells (c), CD3 − CD56 + NK cells (d), γδ and αβ T lymphocytes (e) as well as Vδ1 and Vδ2 subsets (f), present after Ficoll-Paque gradient in gated lympho-mononuclear cells. Panels C and D. CD14 + monocytes (a), CD19 + B cells (b) CD3 − CD56 + NK cells (c), γδ and αβ T lymphocytes (d) as well as Vδ1 and Vδ2 subsets (e), gated mononuclear cells at the end of expansion (day 14) (Panel C) and harvested after αβ T-cell depletion (Panel D). Panel E shows γδ T cell differentiation, by APC-CD27 and PEVio770-CD45RA staining.

Article Snippet: Immunophenotype was assessed on at least 50,000 cells by flow cytometry using fluorochrome-conjugated monoclonal antibodies (mAb): FITC-CD3, PerCP-CD14, Viogreen-CD19, PEVio770 anti-TCRαβ, APC anti-TCRγδ, Viogreen-TCRVδ1, Vioblue-TCRVδ2, and APC-CD56 (all from Miltenyi).

Techniques: Cell Differentiation, Staining