cd34 selection beads kit  (Miltenyi Biotec)

Journal: Cell reports

Article Title: Differentiation latency and dormancy signatures define fetal liver hematopoietic stem cells at single-cell resolution

doi: 10.1016/j.celrep.2025.116289

Figure Lengend Snippet: (A) Sorting strategy for SE hi FL-HSCs from freshly isolated E13.5 FL used for scRNA-seq. (B) Unsupervised clustering of E13.5 SE hi FL-HSC scRNA-seq data in UMAP. (C) Heatmap of gene-set scores by cluster for genes associated with adult HSC dormancy, serial-engrafting HSCs, diapause, , chemokine signaling (WP_CHEMOKINE_SIGNALING_PATHWAY), and a common stem-cell dormancy state associated with lipid metabolism, or genes associated with activated HSC/MPP states including high output and multilineage signatures, , , Myc pathway activation (Hallmark Myc Target Genes V1, V2), and metabolic activity (WP_TCA_CYCLE, HALLMARK_OXIDATIVE_PHOSPHORYLATION, WP_PURINE_METABOLISM). (D) Gene-set expression heatmaps for dormant HSC signature genes and serial-engrafting HSC signature genes. , (E) Expression heatmaps for modules of co-regulated genes. Gene modules 1, 3, 4, and 7 are shown with representative genes identified in each module.

Article Snippet: After centrifugation, the pellet was resuspended in 4 mL MACs buffer (MACS BSA Stock Solution Miltenyi Biotec 130–091-376, in autoMACs Rinsing Solution, Miltenyi Biotec 130–091-222) for CD34 enrichment using CD34 selection beads/kit (Miltenyi Biotech, 130–100-453) according to the manufacturer protocol.

Techniques: Isolation, Activation Assay, Activity Assay, Phospho-proteomics, Expressing

Journal: bioRxiv

Article Title: Generation of effector CD4+ T cells from Human iPSC

doi: 10.1101/2025.08.01.667959

Figure Lengend Snippet: ( A ) Schematic of iT differentiation to CD4+ cells. ( B ) Representative flow cytometry of key developmental timepoints and metrics of DP T cells. iPSCs were plated at day −2 and cultured for 12 days in HSPC differentiation conditions. The resultant CD34+ HSPCs were purified using magnetic beads. Day 12 HSPCs were moved to Notch-stimulatory lymphoid coating and cultured with StemCell Technologies lymphoid expansion media until day 26, where cells were assayed for CD7 expression. Day 26 cells were replated on fresh Notch stimulatory coating and cultured with StemCell Technologies lymphoid maturation media to day 40, where cells are assayed for TCR, CD3, CD4 and CD8 expression. (N=3) ( C ) Representative flow cytometry of D54 T cells, with parent gating in Figure S3A. Day 40 iPSC DP T cells were moved to Retronectin coating and treated with anti-CD3 antibody (OKT3, 5 ug/ml), half media changes were performed every 3 or 4 days. (N=3) ( D ) Representative flow cytometry of D68 T cells post expansion. Day 54 iCD4+ T cells were moved to fresh Retronectin coated wells and treated with media supplemented with anti-CD3/CD28 and 200 U/ml IL-2. (N=3) ( E ) Percentage of CD4 and CD8 positive T cells from iCD4+ T cell differentiation by flow cytometry at day 40, 54 and 68, N=3, analyzed by 2-way ANOVA followed by a Tukey’s multiple comparison test, *P < 0.05; **P < 0.01; ***P < 0.001;****P < 0.0001. ( F ) Representative flow cytometry of T cells from long-term culture in X-Vivo 15 media supplemented with 200 U/ml IL-2. N=3, parent gating for each plot as indicated. (G) Number of doublings of iCD4+ T cells after stimulation with anti-CD3/CD28 (N = 3).

Article Snippet: CD34+ progenitors are isolated from all floating cells by MACS separation using Miltenyi CD34+ selection kit and LS columns (Miltenyi).

Techniques: Flow Cytometry, Cell Culture, Purification, Magnetic Beads, Expressing, Cell Differentiation, Comparison