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B cell development in ABOs is not driven by expansion of pre-existing B-lineage-committed <t>progenitor</t> <t>cells</t> (A) Flow cytometric analysis of <t>CD34,</t> 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 <t>CD34</t> + 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 .
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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