anti human fcr blocking reagent  (Miltenyi Biotec)

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

Journal: Nucleic Acids Research

Article Title: ZEB1 drives terminal erythroid maturation by controlling the GATA2–KLF1 regulatory switch

doi: 10.1093/nar/gkag613

Figure Lengend Snippet: ZEB1 is required for optimal TED in primary human cells. ( A ) Experimental scheme of ZEB1 KD in primary human CD34 + HSPCs. ( B ) ZEB1 mRNA levels measured at day 3 of differentiation. ( C ) GPA levels at day 3 of erythroid differentiation. Percentages of GPA + cells are indicated. ( D ) Cell growth rates at days 3, 7, and 10. ( E ) Proportion of erythroid progenitors, pro-erythroblasts (Pro-E), and basophilic erythroblasts (Baso) at day 3 of erythroid differentiation. ( F ) mRNA levels of GATA2 and KLF1 in ZEB1 KD cells (day 3). ( G ) Enucleation rates measured by Syto-60 staining at day 10 of differentiation. Data are mean ± SEM of n = 3–5 independent biological experiments, from three independent blood donors, * P < .05, ** P < .01, *** P < .001, **** P < .0001 (Student’s t -test).

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 [ ].

Techniques: Staining