macs human cd34 microbead kit  (Miltenyi Biotec)

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

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

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