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cd34 microbead kit  (Miltenyi Biotec)


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    Miltenyi Biotec cd34 microbead kit
    Bone marrow progenitor chimerism 14 months post-allograft. Peripheral blood DNA from both the donor and the recipient, collected before the allograft, were genotyped with KMR Type (DenDx ® ) and analyzed using KMR Engine software (GenDx ® ) according to the manufacturer’s instructions. The markers KMR040 and KMR043 were specific to the recipient and the donor, respectively, and were therefore selected to genotype the BFU-E and CFU-GM colonies in order to identify the progenitor’s origin. <t>CD34</t> + cells were purified from patient’s bone marrow using the <t>CD34</t> MicroBead Kit (human) and plated at a concentration of 250 cells/mL in a methylcellulose-based medium with recombinant cytokines (GFH4435, <t>Stem</t> <t>Cell</t> Technologies) under conditions supporting erythroid and granulocyte/monocyte differentiation. We obtained 44 BFU-E colonies and 24 CFU-GM from 1,500 <t>CD34</t> + HSPCs. After 14 days, colonies were scored and collected individually (24 individual BFU-E and CFU-GM colonies, respectively). Individual colonies were genotyped by qPCR using KMRtype (GenDx). For each colony, both markers were tested alongside an internal control. Colonies that had insufficient internal control or displayed a mixed profile, with detection of both donor and recipient markers (Ct<40), were excluded from the analysis. The qPCR results were interpretable for 17 out of 24 BFU-Es and 5 out of 8 CFU-GMs tested.
    Cd34 Microbead Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1651 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd34+microbead+kit/CD34+MicroBead+Kit%2C+human/pmc13649441-51-48-51
    Average 97 stars, based on 1651 article reviews
    cd34 microbead kit - by Bioz Stars, 2026-10
    97/100 stars

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    1) Product Images from "Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment"

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2026.1932283

    Bone marrow progenitor chimerism 14 months post-allograft. Peripheral blood DNA from both the donor and the recipient, collected before the allograft, were genotyped with KMR Type (DenDx ® ) and analyzed using KMR Engine software (GenDx ® ) according to the manufacturer’s instructions. The markers KMR040 and KMR043 were specific to the recipient and the donor, respectively, and were therefore selected to genotype the BFU-E and CFU-GM colonies in order to identify the progenitor’s origin. CD34 + cells were purified from patient’s bone marrow using the CD34 MicroBead Kit (human) and plated at a concentration of 250 cells/mL in a methylcellulose-based medium with recombinant cytokines (GFH4435, Stem Cell Technologies) under conditions supporting erythroid and granulocyte/monocyte differentiation. We obtained 44 BFU-E colonies and 24 CFU-GM from 1,500 CD34 + HSPCs. After 14 days, colonies were scored and collected individually (24 individual BFU-E and CFU-GM colonies, respectively). Individual colonies were genotyped by qPCR using KMRtype (GenDx). For each colony, both markers were tested alongside an internal control. Colonies that had insufficient internal control or displayed a mixed profile, with detection of both donor and recipient markers (Ct<40), were excluded from the analysis. The qPCR results were interpretable for 17 out of 24 BFU-Es and 5 out of 8 CFU-GMs tested.
    Figure Legend Snippet: Bone marrow progenitor chimerism 14 months post-allograft. Peripheral blood DNA from both the donor and the recipient, collected before the allograft, were genotyped with KMR Type (DenDx ® ) and analyzed using KMR Engine software (GenDx ® ) according to the manufacturer’s instructions. The markers KMR040 and KMR043 were specific to the recipient and the donor, respectively, and were therefore selected to genotype the BFU-E and CFU-GM colonies in order to identify the progenitor’s origin. CD34 + cells were purified from patient’s bone marrow using the CD34 MicroBead Kit (human) and plated at a concentration of 250 cells/mL in a methylcellulose-based medium with recombinant cytokines (GFH4435, Stem Cell Technologies) under conditions supporting erythroid and granulocyte/monocyte differentiation. We obtained 44 BFU-E colonies and 24 CFU-GM from 1,500 CD34 + HSPCs. After 14 days, colonies were scored and collected individually (24 individual BFU-E and CFU-GM colonies, respectively). Individual colonies were genotyped by qPCR using KMRtype (GenDx). For each colony, both markers were tested alongside an internal control. Colonies that had insufficient internal control or displayed a mixed profile, with detection of both donor and recipient markers (Ct<40), were excluded from the analysis. The qPCR results were interpretable for 17 out of 24 BFU-Es and 5 out of 8 CFU-GMs tested.

    Techniques Used: Software, Purification, Concentration Assay, Recombinant, Stem Cell, Control, Analysis

    Related Articles

    Isolation:

    Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch
    Article Snippet: .. CD34 + were isolated by positive selection using the CD34 MicroBead Kit (MACS Miltenyi) according to the manufacturers’ recommendation and cultured following a human ex vivo differentiation protocol as previously described [ ]. .. pTLCV2-sgRNA-Zeb1-up-exon4 (targeting Zeb1 intron 3, and containing a Cas9 expression cassette) and pLentiGuide-Hygro-sgZeb1-downEx6 (targeting Zeb1 intron 6) viral plasmids were generated by inserting annealed sgRNA primers (Table ) into pTLCV2 (Addgene #87360) and pLentiGuide-Hygro (Addgene #139462), respectively.

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Article Title: Temporal Lysophosphatidic Acid Supplementation Enhances Megakaryocyte Differentiation and Platelet Production From Human Hematopoietic Progenitors
    Article Snippet: Umbilical cord blood was diluted with phosphate buffer saline (PBS) at a 1:1 ratio, and gently layered onto Lymphoprep TM (STEMCELL Technologies, Canada), and then centrifuged at 2,000 rpm for 20 min with break. .. Mononuclear cells located between plasma and Lymphoprep TM layers were harvested, washed twice with PBS, and centrifuged at 1,500 rpm for 5 min. HSPCs were isolated from mononuclear cells using a CD34 MicroBead Kit (Miltenyi Biotec, Germany). ..

    Article Title: Cytosolic EZH2-IMPDH2 complexes regulate melanoma progression and metastasis via GTP.
    Article Snippet: Cells from a leukapheresis sample were isolated using Ficoll-Paque PLUS (GE Healthcare) and density centrifugation, followed by NH4Cl lysis to remove red blood cells. .. A secondary isolation step was completed using CD34 MicroBead Kit (Miltenyi Biotec) performed according to the manufacturer’s protocol for positive selection of CD34+ cells from the mononuclear population. .. Isolated CD34+ cells were cultured in expansion medium (Stemspan SFEM (Stem Cell Technologies 09650), 50ng/mL rhFLt3L (R&D 308-FKN), 50ng/mL rhSCF (R&D 255-SC), 10ng/mL rhIL-3 (R&D 203-IL), 10ng/mL rhIL-6 (R&D 206-IL), 35nM UM171 and 500nM Stemreginin) with or without SA for 4 and 7 days.

    Article Title: Wild-Type p53 Protein Enhances APR-246-Induced Cytotoxicity in Acute Myeloid Leukemia and Normal Hematopoietic Stem/Progenitor Cells
    Article Snippet: .. CD34 + cells were isolated using the CD34 Microbead Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). .. Briefly, for positive selection, 1 × 10 8 cells were resuspended in 300 μL of 10% FBS and incubated with 100 μL of FcR blocking buffer and 100 μL of CD34 microbeads (Miltenyi Biotec, 130-100-453) for 30 min in the refrigerator at 4 °C.

    Selection:

    Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch
    Article Snippet: .. CD34 + were isolated by positive selection using the CD34 MicroBead Kit (MACS Miltenyi) according to the manufacturers’ recommendation and cultured following a human ex vivo differentiation protocol as previously described [ ]. .. pTLCV2-sgRNA-Zeb1-up-exon4 (targeting Zeb1 intron 3, and containing a Cas9 expression cassette) and pLentiGuide-Hygro-sgZeb1-downEx6 (targeting Zeb1 intron 6) viral plasmids were generated by inserting annealed sgRNA primers (Table ) into pTLCV2 (Addgene #87360) and pLentiGuide-Hygro (Addgene #139462), respectively.

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Article Title: Cytosolic EZH2-IMPDH2 complexes regulate melanoma progression and metastasis via GTP.
    Article Snippet: Cells from a leukapheresis sample were isolated using Ficoll-Paque PLUS (GE Healthcare) and density centrifugation, followed by NH4Cl lysis to remove red blood cells. .. A secondary isolation step was completed using CD34 MicroBead Kit (Miltenyi Biotec) performed according to the manufacturer’s protocol for positive selection of CD34+ cells from the mononuclear population. .. Isolated CD34+ cells were cultured in expansion medium (Stemspan SFEM (Stem Cell Technologies 09650), 50ng/mL rhFLt3L (R&D 308-FKN), 50ng/mL rhSCF (R&D 255-SC), 10ng/mL rhIL-3 (R&D 203-IL), 10ng/mL rhIL-6 (R&D 206-IL), 35nM UM171 and 500nM Stemreginin) with or without SA for 4 and 7 days.

    Magnetic Cell Separation:

    Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch
    Article Snippet: .. CD34 + were isolated by positive selection using the CD34 MicroBead Kit (MACS Miltenyi) according to the manufacturers’ recommendation and cultured following a human ex vivo differentiation protocol as previously described [ ]. .. pTLCV2-sgRNA-Zeb1-up-exon4 (targeting Zeb1 intron 3, and containing a Cas9 expression cassette) and pLentiGuide-Hygro-sgZeb1-downEx6 (targeting Zeb1 intron 6) viral plasmids were generated by inserting annealed sgRNA primers (Table ) into pTLCV2 (Addgene #87360) and pLentiGuide-Hygro (Addgene #139462), respectively.

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Cell Culture:

    Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch
    Article Snippet: .. CD34 + were isolated by positive selection using the CD34 MicroBead Kit (MACS Miltenyi) according to the manufacturers’ recommendation and cultured following a human ex vivo differentiation protocol as previously described [ ]. .. pTLCV2-sgRNA-Zeb1-up-exon4 (targeting Zeb1 intron 3, and containing a Cas9 expression cassette) and pLentiGuide-Hygro-sgZeb1-downEx6 (targeting Zeb1 intron 6) viral plasmids were generated by inserting annealed sgRNA primers (Table ) into pTLCV2 (Addgene #87360) and pLentiGuide-Hygro (Addgene #139462), respectively.

    Article Title: Foxs1-mediated transformation of CD34 + fibroblast to myCAFs promotes tumor growth.
    Article Snippet: .. These cells were purified with a Cd34 MicroBead Kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and cultured in S/P cell culture medium. ..

    Ex Vivo:

    Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch
    Article Snippet: .. CD34 + were isolated by positive selection using the CD34 MicroBead Kit (MACS Miltenyi) according to the manufacturers’ recommendation and cultured following a human ex vivo differentiation protocol as previously described [ ]. .. pTLCV2-sgRNA-Zeb1-up-exon4 (targeting Zeb1 intron 3, and containing a Cas9 expression cassette) and pLentiGuide-Hygro-sgZeb1-downEx6 (targeting Zeb1 intron 6) viral plasmids were generated by inserting annealed sgRNA primers (Table ) into pTLCV2 (Addgene #87360) and pLentiGuide-Hygro (Addgene #139462), respectively.

    Gradient Centrifugation:

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Purification:

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Article Title: Foxs1-mediated transformation of CD34 + fibroblast to myCAFs promotes tumor growth.
    Article Snippet: .. These cells were purified with a Cd34 MicroBead Kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and cultured in S/P cell culture medium. ..

    Immunostaining:

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment
    Article Snippet: .. Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec). .. Purified CD34+ HSPCs were plated at a concentration of 250 cells/mL in a methylcellulose-based medium (GFH4435, Stem Cell Technologies) under conditions supporting the growth and differentiation of committed erythroid (Burst-Forming Unit-Erythrocytes, BFU-E) and granulo-monocytic (Colony-Forming Unit-Granulocytes Macrophages, CFU-GM) progenitors and incubated at 37 °C in a humidified atmosphere with 5% CO2.

    Clinical Proteomics:

    Article Title: Temporal Lysophosphatidic Acid Supplementation Enhances Megakaryocyte Differentiation and Platelet Production From Human Hematopoietic Progenitors
    Article Snippet: Umbilical cord blood was diluted with phosphate buffer saline (PBS) at a 1:1 ratio, and gently layered onto Lymphoprep TM (STEMCELL Technologies, Canada), and then centrifuged at 2,000 rpm for 20 min with break. .. Mononuclear cells located between plasma and Lymphoprep TM layers were harvested, washed twice with PBS, and centrifuged at 1,500 rpm for 5 min. HSPCs were isolated from mononuclear cells using a CD34 MicroBead Kit (Miltenyi Biotec, Germany). ..



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    Miltenyi Biotec human cd34 microbead kit ultrapure
    NOX1i in Myeloid-HIS-HCC mice reduces monocyte and tumor-associated macrophage infiltration. (A) Schematic overview of the experimental set-up. Male NSG-SGM3 mice (n = 11) were sublethally irradiated and the immune system was partly reconstituted using human cord blood (CB)-derived hematopoietic stem and <t>progenitor</t> <t>cells</t> (HSPCs). Four weeks post humanization, orthotopic human HCC was induced and mice were treated with 50 µM ML171 (NOX1i; n = 6) or vehicle (n = 5), twice per week for 3 weeks. (B) Human-to-murine immune cell (CD45) chimerism in the indicated tissues. (C) Human engraftment based on the number of human CD45 cells. Data are represented as violin plots showing median and quartiles. (D,E) Human immune cell subsets (D) and monocyte subsets (E) in indicated tissues represented as percentage of human CD45 cells. (F) Number of human macrophages and monocytes in the indicated tissues. (G) Representative immunohistochemistry images of infiltrated human monocytes/macrophages in liver tissue of HCC-bearing humanized mice. Sections were stained for human CD14 (brown), or matched IgG control. Scale bars: 100 µm. (B,D,E) . Data are shown as mean (SD). (C–E) Adjusted p-values were calculated using multiple unpaired t-test with Holm-Šídák correction (D,E) , or p-values were calculated using unpaired t-test with Welch’s correction or unpaired Mann-Whitney U test based on normal distribution (C,E) . *p < 0.05. HCC: hepatocellular carcinoma, NOX1i: NOX1 inhibition, huCD45: human CD45, muCD45: murine CD45, NK = natural killer, DCs: dendritic cells.
    Human Cd34 Microbead Kit Ultrapure, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti human fcr blocking reagent  (Miltenyi Biotec)
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    NOX1i in Myeloid-HIS-HCC mice reduces monocyte and tumor-associated macrophage infiltration. (A) Schematic overview of the experimental set-up. Male NSG-SGM3 mice (n = 11) were sublethally irradiated and the immune system was partly reconstituted using human cord blood (CB)-derived hematopoietic stem and <t>progenitor</t> <t>cells</t> (HSPCs). Four weeks post humanization, orthotopic human HCC was induced and mice were treated with 50 µM ML171 (NOX1i; n = 6) or vehicle (n = 5), twice per week for 3 weeks. (B) Human-to-murine immune cell (CD45) chimerism in the indicated tissues. (C) Human engraftment based on the number of human CD45 cells. Data are represented as violin plots showing median and quartiles. (D,E) Human immune cell subsets (D) and monocyte subsets (E) in indicated tissues represented as percentage of human CD45 cells. (F) Number of human macrophages and monocytes in the indicated tissues. (G) Representative immunohistochemistry images of infiltrated human monocytes/macrophages in liver tissue of HCC-bearing humanized mice. Sections were stained for human CD14 (brown), or matched IgG control. Scale bars: 100 µm. (B,D,E) . Data are shown as mean (SD). (C–E) Adjusted p-values were calculated using multiple unpaired t-test with Holm-Šídák correction (D,E) , or p-values were calculated using unpaired t-test with Welch’s correction or unpaired Mann-Whitney U test based on normal distribution (C,E) . *p < 0.05. HCC: hepatocellular carcinoma, NOX1i: NOX1 inhibition, huCD45: human CD45, muCD45: murine CD45, NK = natural killer, DCs: dendritic cells.
    Anti Human Fcr Blocking Reagent, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Bone marrow progenitor chimerism 14 months post-allograft. Peripheral blood DNA from both the donor and the recipient, collected before the allograft, were genotyped with KMR Type (DenDx ® ) and analyzed using KMR Engine software (GenDx ® ) according to the manufacturer’s instructions. The markers KMR040 and KMR043 were specific to the recipient and the donor, respectively, and were therefore selected to genotype the BFU-E and CFU-GM colonies in order to identify the progenitor’s origin. CD34 + cells were purified from patient’s bone marrow using the CD34 MicroBead Kit (human) and plated at a concentration of 250 cells/mL in a methylcellulose-based medium with recombinant cytokines (GFH4435, Stem Cell Technologies) under conditions supporting erythroid and granulocyte/monocyte differentiation. We obtained 44 BFU-E colonies and 24 CFU-GM from 1,500 CD34 + HSPCs. After 14 days, colonies were scored and collected individually (24 individual BFU-E and CFU-GM colonies, respectively). Individual colonies were genotyped by qPCR using KMRtype (GenDx). For each colony, both markers were tested alongside an internal control. Colonies that had insufficient internal control or displayed a mixed profile, with detection of both donor and recipient markers (Ct<40), were excluded from the analysis. The qPCR results were interpretable for 17 out of 24 BFU-Es and 5 out of 8 CFU-GMs tested.

    Journal: Frontiers in Immunology

    Article Title: Case Report: Recurrence of recipient-derived erythropoiesis in hematopoietic stem cell transplant for sickle-cell anemia: multi-lineage chimerism analysis as a tool to distinguish atypical pure red cell aplasia from poor erythroid engraftment

    doi: 10.3389/fimmu.2026.1932283

    Figure Lengend Snippet: Bone marrow progenitor chimerism 14 months post-allograft. Peripheral blood DNA from both the donor and the recipient, collected before the allograft, were genotyped with KMR Type (DenDx ® ) and analyzed using KMR Engine software (GenDx ® ) according to the manufacturer’s instructions. The markers KMR040 and KMR043 were specific to the recipient and the donor, respectively, and were therefore selected to genotype the BFU-E and CFU-GM colonies in order to identify the progenitor’s origin. CD34 + cells were purified from patient’s bone marrow using the CD34 MicroBead Kit (human) and plated at a concentration of 250 cells/mL in a methylcellulose-based medium with recombinant cytokines (GFH4435, Stem Cell Technologies) under conditions supporting erythroid and granulocyte/monocyte differentiation. We obtained 44 BFU-E colonies and 24 CFU-GM from 1,500 CD34 + HSPCs. After 14 days, colonies were scored and collected individually (24 individual BFU-E and CFU-GM colonies, respectively). Individual colonies were genotyped by qPCR using KMRtype (GenDx). For each colony, both markers were tested alongside an internal control. Colonies that had insufficient internal control or displayed a mixed profile, with detection of both donor and recipient markers (Ct<40), were excluded from the analysis. The qPCR results were interpretable for 17 out of 24 BFU-Es and 5 out of 8 CFU-GMs tested.

    Article Snippet: Bone marrow mononuclear cells were first isolated by density gradient centrifugation over Lymphocytes Separation Medium (MSL , Eurobio Scientific; density 1.078 g/mL), and CD34+ multipotent hematopoietic stem/progenitor cells (HSPCs) were subsequently purified from the mononuclear fraction by immunomagnetic selection with MACS columns (Miltenyi Biotec) after immunostaining with the CD34 MicroBead Kit (Miltenyi Biotec).

    Techniques: Software, Purification, Concentration Assay, Recombinant, Stem Cell, Control, Analysis

    B cell development in ABOs is not driven by expansion of pre-existing B-lineage-committed progenitor cells (A) Flow cytometric analysis of CD34, c-KIT (CD117), CD38, and CD10 expression on HSPCs before (top) and after depletion of Lin + CD10 + cells (bottom) by MACS, visualized with opt-SNE. (B and C) Flow cytometric analysis comparing the frequencies of CD33 + myeloid-committed (B) and CD10 + CD38 + lymphoid-committed cells (C) in day 35 ABOs using bulk or Lin + CD10 + -depleted CD34 + HSPCs. (D) Quantification of CD33 + and CD10 + CD38 + cells in day 35 ABOs. (E) Flow cytometric analysis comparing IgM + IgD − (immature) and IgM + IgD + (transitional) B cell frequencies in day 35 ABOs using bulk or Lin + CD10 + -depleted CD34 + HSPCs. (F) Quantification of IgM + and IgM + IgD + B cell frequencies in day 35 ABOs. (G) Distribution of HSPCs along the B cell developmental trajectory in day 35 ABOs. (D, F, G) Data are presented as mean ± SEM. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s multiple comparisons test (ns; non-significant). (A–G) Data represent n = 3 individual donors in independent cultures. See also .

    Journal: Cell Reports Medicine

    Article Title: ABO: A 3D stroma-supported culture platform enabling full human B-lymphopoiesis for disease modeling and gene therapy development

    doi: 10.1016/j.xcrm.2026.102879

    Figure Lengend Snippet: B cell development in ABOs is not driven by expansion of pre-existing B-lineage-committed progenitor cells (A) Flow cytometric analysis of CD34, c-KIT (CD117), CD38, and CD10 expression on HSPCs before (top) and after depletion of Lin + CD10 + cells (bottom) by MACS, visualized with opt-SNE. (B and C) Flow cytometric analysis comparing the frequencies of CD33 + myeloid-committed (B) and CD10 + CD38 + lymphoid-committed cells (C) in day 35 ABOs using bulk or Lin + CD10 + -depleted CD34 + HSPCs. (D) Quantification of CD33 + and CD10 + CD38 + cells in day 35 ABOs. (E) Flow cytometric analysis comparing IgM + IgD − (immature) and IgM + IgD + (transitional) B cell frequencies in day 35 ABOs using bulk or Lin + CD10 + -depleted CD34 + HSPCs. (F) Quantification of IgM + and IgM + IgD + B cell frequencies in day 35 ABOs. (G) Distribution of HSPCs along the B cell developmental trajectory in day 35 ABOs. (D, F, G) Data are presented as mean ± SEM. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s multiple comparisons test (ns; non-significant). (A–G) Data represent n = 3 individual donors in independent cultures. See also .

    Article Snippet: Leukocytes from UCB and mPB were first purified using the Erythrocyte Sedimentation Kit II (Miltenyi), followed by CD34 + cell enrichment with the CD34 MicroBead Kit UltraPure (Miltenyi) per manufacturer’s instructions.

    Techniques: Expressing

    NOX1i in Myeloid-HIS-HCC mice reduces monocyte and tumor-associated macrophage infiltration. (A) Schematic overview of the experimental set-up. Male NSG-SGM3 mice (n = 11) were sublethally irradiated and the immune system was partly reconstituted using human cord blood (CB)-derived hematopoietic stem and progenitor cells (HSPCs). Four weeks post humanization, orthotopic human HCC was induced and mice were treated with 50 µM ML171 (NOX1i; n = 6) or vehicle (n = 5), twice per week for 3 weeks. (B) Human-to-murine immune cell (CD45) chimerism in the indicated tissues. (C) Human engraftment based on the number of human CD45 cells. Data are represented as violin plots showing median and quartiles. (D,E) Human immune cell subsets (D) and monocyte subsets (E) in indicated tissues represented as percentage of human CD45 cells. (F) Number of human macrophages and monocytes in the indicated tissues. (G) Representative immunohistochemistry images of infiltrated human monocytes/macrophages in liver tissue of HCC-bearing humanized mice. Sections were stained for human CD14 (brown), or matched IgG control. Scale bars: 100 µm. (B,D,E) . Data are shown as mean (SD). (C–E) Adjusted p-values were calculated using multiple unpaired t-test with Holm-Šídák correction (D,E) , or p-values were calculated using unpaired t-test with Welch’s correction or unpaired Mann-Whitney U test based on normal distribution (C,E) . *p < 0.05. HCC: hepatocellular carcinoma, NOX1i: NOX1 inhibition, huCD45: human CD45, muCD45: murine CD45, NK = natural killer, DCs: dendritic cells.

    Journal: Frontiers in Pharmacology

    Article Title: Harnessing human immune system models to validate NADPH oxidase 1 inhibition as treatment for hepatocellular carcinoma

    doi: 10.3389/fphar.2026.1808515

    Figure Lengend Snippet: NOX1i in Myeloid-HIS-HCC mice reduces monocyte and tumor-associated macrophage infiltration. (A) Schematic overview of the experimental set-up. Male NSG-SGM3 mice (n = 11) were sublethally irradiated and the immune system was partly reconstituted using human cord blood (CB)-derived hematopoietic stem and progenitor cells (HSPCs). Four weeks post humanization, orthotopic human HCC was induced and mice were treated with 50 µM ML171 (NOX1i; n = 6) or vehicle (n = 5), twice per week for 3 weeks. (B) Human-to-murine immune cell (CD45) chimerism in the indicated tissues. (C) Human engraftment based on the number of human CD45 cells. Data are represented as violin plots showing median and quartiles. (D,E) Human immune cell subsets (D) and monocyte subsets (E) in indicated tissues represented as percentage of human CD45 cells. (F) Number of human macrophages and monocytes in the indicated tissues. (G) Representative immunohistochemistry images of infiltrated human monocytes/macrophages in liver tissue of HCC-bearing humanized mice. Sections were stained for human CD14 (brown), or matched IgG control. Scale bars: 100 µm. (B,D,E) . Data are shown as mean (SD). (C–E) Adjusted p-values were calculated using multiple unpaired t-test with Holm-Šídák correction (D,E) , or p-values were calculated using unpaired t-test with Welch’s correction or unpaired Mann-Whitney U test based on normal distribution (C,E) . *p < 0.05. HCC: hepatocellular carcinoma, NOX1i: NOX1 inhibition, huCD45: human CD45, muCD45: murine CD45, NK = natural killer, DCs: dendritic cells.

    Article Snippet: Freshly isolated CB-derived PBMCs, isolated from CB as described for buffy coat, were enriched for HSPCs based on positive immunomagnetic selection of CD34 + expressing cells using the human CD34 MicroBead Kit UltraPure (Miltenyi Biotec, #130–100-453) according to manufacturer’s guidelines.

    Techniques: Irradiation, Derivative Assay, Immunohistochemistry, Staining, Control, MANN-WHITNEY, Inhibition