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cd34 cd38 cell isolation kit  (Miltenyi Biotec)


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    Miltenyi Biotec cd34 cd38 cell isolation kit
    Cd34 Cd38 Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd34+cd38+cell+isolation+kit/CD34%2BCD38-+Cell+Isolation+Kit%2C+human/pm41809821-64-12-18
    Average 95 stars, based on 3 article reviews
    cd34 cd38 cell isolation kit - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Isolation:

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells.
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1a cells and primary AML cells by CD34+CD38– Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1a cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL-15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1α cells and primary AML cells by CD34 + CD38 – Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1α cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Cell Isolation:

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells.
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1a cells and primary AML cells by CD34+CD38– Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1a cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Article Title: Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol
    Article Snippet: .. Cord blood CD34+38- were purified by means of CD34+CD38- Cell Isolation Kit, human (Miltenyi Biotec; Bergisch Gladbach, Germany). .. Cell purity was assessed by means of flow cytometry after immunostaining with anti-human CD34-APC (1:50) and Labeling Check Reagent-PE (1:33), Labeling Check Reagent-FITC (1:33) or antibodies (Miltenyi Biotec).

    Article Title: Spred1 deficit promotes treatment resistance and transformation of chronic phase CML
    Article Snippet: .. Flow cytometry analyses Human CD34 + cells were selected using the indirect CD34 microbead kit (Miltenyi Biotec, San Diego, CA) and CD34 + CD38 − cells were sorted after staining with human antibodies against CD34 and CD38 ( Supplementary Table 1 ) or selected using CD34+CD38- cell isolation kit (Miltenyi Biotec, San Diego, CA) according to the manufacturer’s protocol. ..

    Article Title: Targeting HSP90 dimerization via the C terminus is effective in imatinib-resistant CML and lacks the heat shock response
    Article Snippet: Primary patient samples were obtained from newly diagnosed or relapsed patients (supplemental Table 2) after informed consent approval of the local ethics committee and were cultured either in Stemline II Hematopoietic Stem Cell Expansion Medium (Sigma-Aldrich) or in Mononuclear Cell Medium (PromoCell). .. CML and BCR-ABL1+ BCP-ALL samples were sorted for CD34 + CD38 +/− using the CD34 + CD38 − Cell Isolation Kit, human (Miltenyi Biotec). .. Inhibitors were printed on white 96-well or 384-well plates (Thermo Fisher Scientific) with their increasing concentration (50 nM to 25 μM) along with respective controls by using a digital dispenser (D300e; Tecan, Männedorf, Switzerland).

    Article Title: Spred1 deficit promotes treatment resistance and transformation of chronic phase CML
    Article Snippet: .. Human CD34 + cells were selected using the indirect CD34 microbead kit (Miltenyi Biotec, San Diego, CA) and CD34 + CD38 − cells were sorted after staining with human antibodies against CD34 and CD38 ( Supplementary Table 1 ) or selected using CD34+CD38- cell isolation kit (Miltenyi Biotec, San Diego, CA) according to the manufacturer’s protocol. ..

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL-15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1α cells and primary AML cells by CD34 + CD38 – Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1α cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Magnetic Cell Separation:

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells.
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1a cells and primary AML cells by CD34+CD38– Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1a cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Article Title: cGAS/STING-mediated upregulation of NKG2D ligands in LSCs contributes to enhanced sensitivity to NK cells
    Article Snippet: Primary NK cells were cultured in RPMI-1640 Medium, complemented with 10% FBS (Gibco, Australia), 200 IU/mL rhIL-2 (Beyotime, China), 10 ng/mL rhIL-15 (Beyotime, China) and 1% penicillin and streptomycin. .. The LSCs were isolated from KG-1α cells and primary AML cells by CD34 + CD38 – Cell Isolation Kit (Cat#: 130-114-822, Miltenyi Biotec) via MACS Separation as instructions. .. Briefly, KG-1α cells or primary AML cells were collected in cold medium by centrifuge at 300×g for 10 minutes, following by the steps sequentially: First magnetic labeling (incubation with CD34 MultiSort MicroBeads)-First magnetic separation-Removal of MultiSort MicroBeads and second magnetic labeling (incubation with CD34 MultiSort MicroBeads and CD38 MicroBeads)-Second magnetic separation with LS Columns.

    Purification:

    Article Title: Genomic Analysis of Hematopoietic Stem Cell at the Single-Cell Level: Optimization of Cell Fixation and Whole Genome Amplification (WGA) Protocol
    Article Snippet: .. Cord blood CD34+38- were purified by means of CD34+CD38- Cell Isolation Kit, human (Miltenyi Biotec; Bergisch Gladbach, Germany). .. Cell purity was assessed by means of flow cytometry after immunostaining with anti-human CD34-APC (1:50) and Labeling Check Reagent-PE (1:33), Labeling Check Reagent-FITC (1:33) or antibodies (Miltenyi Biotec).

    Flow Cytometry:

    Article Title: Spred1 deficit promotes treatment resistance and transformation of chronic phase CML
    Article Snippet: .. Flow cytometry analyses Human CD34 + cells were selected using the indirect CD34 microbead kit (Miltenyi Biotec, San Diego, CA) and CD34 + CD38 − cells were sorted after staining with human antibodies against CD34 and CD38 ( Supplementary Table 1 ) or selected using CD34+CD38- cell isolation kit (Miltenyi Biotec, San Diego, CA) according to the manufacturer’s protocol. ..

    Staining:

    Article Title: Spred1 deficit promotes treatment resistance and transformation of chronic phase CML
    Article Snippet: .. Flow cytometry analyses Human CD34 + cells were selected using the indirect CD34 microbead kit (Miltenyi Biotec, San Diego, CA) and CD34 + CD38 − cells were sorted after staining with human antibodies against CD34 and CD38 ( Supplementary Table 1 ) or selected using CD34+CD38- cell isolation kit (Miltenyi Biotec, San Diego, CA) according to the manufacturer’s protocol. ..

    Article Title: Spred1 deficit promotes treatment resistance and transformation of chronic phase CML
    Article Snippet: .. Human CD34 + cells were selected using the indirect CD34 microbead kit (Miltenyi Biotec, San Diego, CA) and CD34 + CD38 − cells were sorted after staining with human antibodies against CD34 and CD38 ( Supplementary Table 1 ) or selected using CD34+CD38- cell isolation kit (Miltenyi Biotec, San Diego, CA) according to the manufacturer’s protocol. ..



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    Image Search Results


    a , Schematic of the experimental design for de novo generation of human T-ALL. Sublethally irradiated NSG mice were subjected to i.v. injection with human cord blood (CB) CD34 + HPCs (1–1.5 × 10 5 cells per mouse) retrovirally transduced with either ICN1 along with GFP, or GFP alone as the control. b , Representative flow cytometry expression of CD4, CD8, CD3, TCRαβ and TCRγδ by ICN1 + human T-ALL cells infiltrating the bone marrow of diseased mice in a at 27 weeks after transplant. c , Representative CD3 versus TCRαβ expression of human ICN1 + T-ALL cells recovered from diseased mice in a (primary), or from NSG mice after serial transplantation with bone marrow from primary mice (1st transfer and 2nd transfer). Results correspond to T-ALL cells isolated from the bone marrow at 39, 8 and 9 weeks after transplant, respectively. d , Mean percentages ± s.e.m. of human CD8 + CD4 + double-positive, CD3 lo TCRαβ neg ICN1 + cells infiltrating the bone marrow of either primary diseased mice, or serially transplanted mice (nine mice per group from two independent experiments) as in c . Data were analyzed by one-way analysis of variance (ANOVA) with Kruskal–Wallis Dunn’s multiple-comparisons test. **** P < 0.0001.

    Journal: Nature Immunology

    Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

    doi: 10.1038/s41590-025-02265-w

    Figure Lengend Snippet: a , Schematic of the experimental design for de novo generation of human T-ALL. Sublethally irradiated NSG mice were subjected to i.v. injection with human cord blood (CB) CD34 + HPCs (1–1.5 × 10 5 cells per mouse) retrovirally transduced with either ICN1 along with GFP, or GFP alone as the control. b , Representative flow cytometry expression of CD4, CD8, CD3, TCRαβ and TCRγδ by ICN1 + human T-ALL cells infiltrating the bone marrow of diseased mice in a at 27 weeks after transplant. c , Representative CD3 versus TCRαβ expression of human ICN1 + T-ALL cells recovered from diseased mice in a (primary), or from NSG mice after serial transplantation with bone marrow from primary mice (1st transfer and 2nd transfer). Results correspond to T-ALL cells isolated from the bone marrow at 39, 8 and 9 weeks after transplant, respectively. d , Mean percentages ± s.e.m. of human CD8 + CD4 + double-positive, CD3 lo TCRαβ neg ICN1 + cells infiltrating the bone marrow of either primary diseased mice, or serially transplanted mice (nine mice per group from two independent experiments) as in c . Data were analyzed by one-way analysis of variance (ANOVA) with Kruskal–Wallis Dunn’s multiple-comparisons test. **** P < 0.0001.

    Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

    Techniques: Irradiation, Injection, Transduction, Control, Flow Cytometry, Expressing, Transplantation Assay, Isolation

    ( A ) Representative pseudocolour plots of CD3 vs pTα expression in pre-TCR-expressing SupT1, and TCRαβ-expressing HPB-ALL and Jurkat human cell lines from three independent experiments, as assessed by reactivity with anti-CD3ε and anti-pTα monoclonal antibodies (left biparametric histograms). Representative monoparametric histograms on the right show surface pre-TCR expression levels analysed with anti-pTα monoclonal antibody plus goat anti-mouse FITC. Numbers indicate percentages of positive cells. Shaded histograms show background staining with an irrelevant isotype-matched antibody. ( B ) Representative CD3 vs TCRαβ expression of either primary human thymocytes from a single 17-weeks-old fetal thymus sample (upper left), or developing T cells derived either from two independent human CB HPC samples after 70 days of culture onto OP9-Jag2 cells (middle left), or from a single CD34 + human ETP sample after 18 days of culture in ATOs (bottom left). Pre-TCR expression levels of electronically gated CD3 lo TCRαβ neg , TCRαβ + and CD3 neg cells analysed with anti-pTα mAb plus biotin-coupled anti-mouse (Fab’)2 plus BV421-coupled streptavidin are shown in the monoparametric histograms on the right. Numbers indicate percentages of positive cells relative to background staining with an isotype-matched mAb.

    Journal: Nature Immunology

    Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

    doi: 10.1038/s41590-025-02265-w

    Figure Lengend Snippet: ( A ) Representative pseudocolour plots of CD3 vs pTα expression in pre-TCR-expressing SupT1, and TCRαβ-expressing HPB-ALL and Jurkat human cell lines from three independent experiments, as assessed by reactivity with anti-CD3ε and anti-pTα monoclonal antibodies (left biparametric histograms). Representative monoparametric histograms on the right show surface pre-TCR expression levels analysed with anti-pTα monoclonal antibody plus goat anti-mouse FITC. Numbers indicate percentages of positive cells. Shaded histograms show background staining with an irrelevant isotype-matched antibody. ( B ) Representative CD3 vs TCRαβ expression of either primary human thymocytes from a single 17-weeks-old fetal thymus sample (upper left), or developing T cells derived either from two independent human CB HPC samples after 70 days of culture onto OP9-Jag2 cells (middle left), or from a single CD34 + human ETP sample after 18 days of culture in ATOs (bottom left). Pre-TCR expression levels of electronically gated CD3 lo TCRαβ neg , TCRαβ + and CD3 neg cells analysed with anti-pTα mAb plus biotin-coupled anti-mouse (Fab’)2 plus BV421-coupled streptavidin are shown in the monoparametric histograms on the right. Numbers indicate percentages of positive cells relative to background staining with an isotype-matched mAb.

    Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

    Techniques: Expressing, Bioprocessing, Staining, Derivative Assay

    ( A ) Immunoblot analysis of CMS expression in SupT1 cells that were either transfected with several shRNAs (shCMS1-5) specific for CD2AP , the gene encoding CMS, or left untransfected (NT) (left panel). Relative CMS expression normalized to α-tubulin expression used as loading control is shown in the right panel. ( B ) Immunoblot analysis of CMS expression (upper) and PLCγ tyrosine (Tyr) phosphorylation (bottom) in JR.pTα pre-TCR + cells transduced either with shCMS4 or with shSC as control, prior to activation with an anti-CD3ε monoclonal antibody for the indicated times. PLCγ Tyr-phosphorylation was analysed after immunoprecipitation with anti-PLCγ1 monoclonal antibody by probing with an anti-Y783-PLCγ1 antibody. Tubulin expression was analysed as loading control. ( C ) Relative NFAT activity of JR.pTα cells transduced with either shCMS4 or shSC as control, analysed upon stimulation with an anti-CD3ε monoclonal antibody by a luciferase assay. Data are shown as mean percentages ± SEM of NFAT activity relative to activated shSC cells analysed by two-tailed unpaired t test (n = 3). **** P < 0.0001. ( D ) Immunoblot analysis of CMS in human CD34 + early thymic progenitors (ETPs) transduced with a lentivirus encoding either shCMS4 and GFP or shSC and GFP as control. ( E ) Absolute numbers of shCMS- or shSC-transduced ETP-derived human cells reconstituting the thymus of RAG-2 −/− γc −/− mice at the indicated weeks post-transplant (n = 6 week 3, n = 4 week 5; *** P = 0.0002). ETP transduction efficiencies with shCMS and shSC were 24,5% ± 4,5% and 21,5% ± 3,5%, respectively. ( F ) Absolute numbers of shCMS- or shSC-transduced human thymocytes in ( E ) expressing the double positive CD3 lo TCRαβ − phenotype at 3- and 5-weeks post-transplant (left panel; * P = 0.0222) or the post-β selected DP CD3 + TCRαβ + phenotype at 5-weeks post-transplant (right panel; *P = 0.0146). Data in ( E, F ) are shown as mean numbers ± s.e.m. of transduced (GFP + ) cells normalized to 10 5 transduced input cells from two independent experiments, analysed by two-tailed unpaired t test.

    Journal: Nature Immunology

    Article Title: Pre-TCR-targeted immunotherapy for T cell acute lymphoblastic leukemia

    doi: 10.1038/s41590-025-02265-w

    Figure Lengend Snippet: ( A ) Immunoblot analysis of CMS expression in SupT1 cells that were either transfected with several shRNAs (shCMS1-5) specific for CD2AP , the gene encoding CMS, or left untransfected (NT) (left panel). Relative CMS expression normalized to α-tubulin expression used as loading control is shown in the right panel. ( B ) Immunoblot analysis of CMS expression (upper) and PLCγ tyrosine (Tyr) phosphorylation (bottom) in JR.pTα pre-TCR + cells transduced either with shCMS4 or with shSC as control, prior to activation with an anti-CD3ε monoclonal antibody for the indicated times. PLCγ Tyr-phosphorylation was analysed after immunoprecipitation with anti-PLCγ1 monoclonal antibody by probing with an anti-Y783-PLCγ1 antibody. Tubulin expression was analysed as loading control. ( C ) Relative NFAT activity of JR.pTα cells transduced with either shCMS4 or shSC as control, analysed upon stimulation with an anti-CD3ε monoclonal antibody by a luciferase assay. Data are shown as mean percentages ± SEM of NFAT activity relative to activated shSC cells analysed by two-tailed unpaired t test (n = 3). **** P < 0.0001. ( D ) Immunoblot analysis of CMS in human CD34 + early thymic progenitors (ETPs) transduced with a lentivirus encoding either shCMS4 and GFP or shSC and GFP as control. ( E ) Absolute numbers of shCMS- or shSC-transduced ETP-derived human cells reconstituting the thymus of RAG-2 −/− γc −/− mice at the indicated weeks post-transplant (n = 6 week 3, n = 4 week 5; *** P = 0.0002). ETP transduction efficiencies with shCMS and shSC were 24,5% ± 4,5% and 21,5% ± 3,5%, respectively. ( F ) Absolute numbers of shCMS- or shSC-transduced human thymocytes in ( E ) expressing the double positive CD3 lo TCRαβ − phenotype at 3- and 5-weeks post-transplant (left panel; * P = 0.0222) or the post-β selected DP CD3 + TCRαβ + phenotype at 5-weeks post-transplant (right panel; *P = 0.0146). Data in ( E, F ) are shown as mean numbers ± s.e.m. of transduced (GFP + ) cells normalized to 10 5 transduced input cells from two independent experiments, analysed by two-tailed unpaired t test.

    Article Snippet: HPCs were obtained from Ficoll-Hypaque-purified cord blood samples using the CD34 Progenitor Cell Isolation Kit (Miltenyi Biotec) .

    Techniques: Western Blot, Expressing, Transfection, Control, Phospho-proteomics, Activation Assay, Immunoprecipitation, Activity Assay, Transduction, Luciferase, Two Tailed Test, Derivative Assay

    BMMC samples derived from AML and MDS patients were treated with 0–500 pM of AFM28, a non-targeting control (RSV/CD16A) or an Fc-enhanced anti-CD123 IgG antibody for 24 h in the presence of IL-2-preincubated allogeneic healthy donor NK cells, all derived from different donors, at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + or CD33 + /CD38 + /CD123 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment were set to baseline. A – D Concentration-dependent lysis of blasts from AML ( A , B , n = 10) or MDS ( C , D , n = 5) patients by AFM28 compared to RSV/CD16A. E – H Concentration-dependent lysis of blasts from AML patients with low ( E , F , n = 5) or high ( G , H , n = 5) CD64 MFI by AFM28 compared to an Fc-enhanced anti-CD123 IgG antibody. Data in ( A , C , E and G ) are represented as mean ± SD. Data in ( B , D , F and H ) were analyzed using two-way ANOVA and Šídák’s multiple comparisons test. MFI median fluorescence intensity; SD standard deviation.

    Journal: Nature Communications

    Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

    doi: 10.1038/s41467-025-63069-y

    Figure Lengend Snippet: BMMC samples derived from AML and MDS patients were treated with 0–500 pM of AFM28, a non-targeting control (RSV/CD16A) or an Fc-enhanced anti-CD123 IgG antibody for 24 h in the presence of IL-2-preincubated allogeneic healthy donor NK cells, all derived from different donors, at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + or CD33 + /CD38 + /CD123 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment were set to baseline. A – D Concentration-dependent lysis of blasts from AML ( A , B , n = 10) or MDS ( C , D , n = 5) patients by AFM28 compared to RSV/CD16A. E – H Concentration-dependent lysis of blasts from AML patients with low ( E , F , n = 5) or high ( G , H , n = 5) CD64 MFI by AFM28 compared to an Fc-enhanced anti-CD123 IgG antibody. Data in ( A , C , E and G ) are represented as mean ± SD. Data in ( B , D , F and H ) were analyzed using two-way ANOVA and Šídák’s multiple comparisons test. MFI median fluorescence intensity; SD standard deviation.

    Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

    Techniques: Derivative Assay, Control, Flow Cytometry, Concentration Assay, Lysis, Fluorescence, Standard Deviation

    BMMC patient-derived AML and MDS samples were treated with or without 100 pM of AFM28 for 24 h in the presence of IL-2-preincubated allogeneic NK cells, all derived from different donors, at an E:T ratio of 1:1. Analysis was performed using flow cytometry. LSPCs were defined as viable/CD45 low /CD34 + /CD38 − /CD117 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment for LSPCs and CD123 + LSPCs were set to baseline. A Representative dot plot of LSPC lysis following treatment with 100 pM AFM28 or without AFM28 (indicated as 0 pM) of one AML patient sample. B , C Lysis of LSPCs from AML patients ( B , n = 5) and from MDS patients ( C , n = 5) in the presence of 100 pM AFM28 or without AFM28. Data are represented as mean ± SD and were analyzed using one-way and two-way ANOVA and Šídák’s multiple comparisons test. D , Schematic workflow of the CFU assay. Created in BioRender ( https://BioRender.com/8l2brjy ). E – G CFU assay results of AML ( E , n = 5), MDS ( F , n = 5) and healthy ( G , n = 5) CD34 + cell samples treated with 0/10/100/1000 pM of AFM28 for 24 h in the presence of allogeneic NK cells at an E:T ratio of 1:1. “CD34 + only” describes culturing untreated CD34 + cells without allogeneic NK cells. Colonies were counted manually. Colony count of “CD34 + only” condition was normalized to 100%. Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. SD standard deviation.

    Journal: Nature Communications

    Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

    doi: 10.1038/s41467-025-63069-y

    Figure Lengend Snippet: BMMC patient-derived AML and MDS samples were treated with or without 100 pM of AFM28 for 24 h in the presence of IL-2-preincubated allogeneic NK cells, all derived from different donors, at an E:T ratio of 1:1. Analysis was performed using flow cytometry. LSPCs were defined as viable/CD45 low /CD34 + /CD38 − /CD117 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment for LSPCs and CD123 + LSPCs were set to baseline. A Representative dot plot of LSPC lysis following treatment with 100 pM AFM28 or without AFM28 (indicated as 0 pM) of one AML patient sample. B , C Lysis of LSPCs from AML patients ( B , n = 5) and from MDS patients ( C , n = 5) in the presence of 100 pM AFM28 or without AFM28. Data are represented as mean ± SD and were analyzed using one-way and two-way ANOVA and Šídák’s multiple comparisons test. D , Schematic workflow of the CFU assay. Created in BioRender ( https://BioRender.com/8l2brjy ). E – G CFU assay results of AML ( E , n = 5), MDS ( F , n = 5) and healthy ( G , n = 5) CD34 + cell samples treated with 0/10/100/1000 pM of AFM28 for 24 h in the presence of allogeneic NK cells at an E:T ratio of 1:1. “CD34 + only” describes culturing untreated CD34 + cells without allogeneic NK cells. Colonies were counted manually. Colony count of “CD34 + only” condition was normalized to 100%. Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. SD standard deviation.

    Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

    Techniques: Derivative Assay, Flow Cytometry, Lysis, Colony-forming Unit Assay, Standard Deviation

    A Fresh whole blood samples from newly diagnosed AML patients ( n = 6) were treated with AFM28 (0/0.01/1 µg/ml equivalent to 0/50/5000 pM) or vehicle in the absence (circles) or presence of healthy donor-derived allogeneic NK cells (squares: fresh, non-expanded NK cells; triangles: cryopreserved cytokine-expanded NK cells generated by a standardized protocol, see Supplementary Methods and Supplementary Fig. ), all derived from different donors, for 24 h at an E:T ratio (NK cells:peripheral leukocytes) of 1:1. Blasts were defined as viable/CD45 low /CD123 + /CD33 + /CD117 + or CD34 + /CD117 + . The gating strategy is shown in Supplementary Fig. . Leukemic blast counts of vehicle treatment without NK cells were set to baseline. E:T ratios of patient’s endogenous NK cells to blasts (in the absence of healthy donor-derived allogeneic NK cells) were 0.1:1, 0.01:1 and 0.2:1 for non-responders and 0.5:1, 0.1:1 and 0.1:1 for responders (data points from top to bottom for 5000 pM AFM28). Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. B BMMC samples derived from a single AML patient were treated with 0–500 pM of AFM28 for 24 h in the presence of allogeneic AML patient-derived NK cells ( n = 5) or healthy donor-derived NK cells ( n = 5) at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + /CD38 + /CD123 + . Blast counts of 0 pM treatment were set to baseline. HY healthy, SD standard deviation.

    Journal: Nature Communications

    Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

    doi: 10.1038/s41467-025-63069-y

    Figure Lengend Snippet: A Fresh whole blood samples from newly diagnosed AML patients ( n = 6) were treated with AFM28 (0/0.01/1 µg/ml equivalent to 0/50/5000 pM) or vehicle in the absence (circles) or presence of healthy donor-derived allogeneic NK cells (squares: fresh, non-expanded NK cells; triangles: cryopreserved cytokine-expanded NK cells generated by a standardized protocol, see Supplementary Methods and Supplementary Fig. ), all derived from different donors, for 24 h at an E:T ratio (NK cells:peripheral leukocytes) of 1:1. Blasts were defined as viable/CD45 low /CD123 + /CD33 + /CD117 + or CD34 + /CD117 + . The gating strategy is shown in Supplementary Fig. . Leukemic blast counts of vehicle treatment without NK cells were set to baseline. E:T ratios of patient’s endogenous NK cells to blasts (in the absence of healthy donor-derived allogeneic NK cells) were 0.1:1, 0.01:1 and 0.2:1 for non-responders and 0.5:1, 0.1:1 and 0.1:1 for responders (data points from top to bottom for 5000 pM AFM28). Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. B BMMC samples derived from a single AML patient were treated with 0–500 pM of AFM28 for 24 h in the presence of allogeneic AML patient-derived NK cells ( n = 5) or healthy donor-derived NK cells ( n = 5) at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + /CD38 + /CD123 + . Blast counts of 0 pM treatment were set to baseline. HY healthy, SD standard deviation.

    Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

    Techniques: Derivative Assay, Generated, Flow Cytometry, Standard Deviation