cd34 cd38 cell isolation kit  (Miltenyi Biotec)

Journal: Nature Communications

Article Title: Circadian fluctuation of soluble CD26 dictates the impact of the timing of cord blood transplantation on acute graft-versus-host disease

doi: 10.1038/s41467-026-68958-4

Figure Lengend Snippet: a Diagram of patients who underwent UCBT. Patients were divided into two groups on the basis of stem cell infusion timing, with a cutoff time of 9:40 am. UCBT, unrelated cord blood transplantation; CR, complete remission; MRD minimal residual disease; CB, cord blood. b Histogram showing the distribution of stem cell infusion times in the cohort. c , d Cumulative incidence (CI) of grade II-IV aGVHD ( c ) ( P = 0.416) and grade III-IV aGVHD ( d ) ( P = 0.004) in the earlier infusion (≤ 9:40 am, n = 219) and later infusion (> 9:40 am, n = 215) groups. The P values are two-sided and reported as exact values. e CI of overall chronic GVHD (cGVHD) in the earlier infusion (≤ 9:40 am) and later infusion (> 9:40 am) groups ( P = 0.276). The P values are two-sided and reported as exact values. f CI of 3-year TRM in the earlier infusion (≤ 9:40 am, n = 219) and later infusion (> 9:40 am, n = 215) groups ( P = 0.067). The P values are two-sided and reported as exact values. g – i Probabilities of overall survival ( g ) ( P = 0.022), disease-free survival ( h ) ( P = 0.050), and GRFS ( i ) ( P < 0.001) in the earlier infusion (≤ 9:40 am, n = 219) and later infusion (> 9:40 am, n = 215) groups. j CI of 3-year relapse in the earlier infusion (≤ 9:40 am, n = 219) and later infusion (> 9:40 am, n = 215) groups ( P = 0.775). The P values are two-sided and reported as exact values.The data were analyzed by Gray’s test ( c – f , and j ) and the log-rank test ( g – i ). Source data are provided as a Source Data file.

Article Snippet: For CD34+ cell isolation, immunomagnetic depletion was performed via CD34 microbeads (130-046-702, Miltenyi Biotech) following the recommended protocol.

Techniques: Transplantation Assay

Journal: Nature Communications

Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

doi: 10.1038/s41467-025-63069-y

Figure Lengend Snippet: BMMC samples derived from AML and MDS patients were treated with 0–500 pM of AFM28, a non-targeting control (RSV/CD16A) or an Fc-enhanced anti-CD123 IgG antibody for 24 h in the presence of IL-2-preincubated allogeneic healthy donor NK cells, all derived from different donors, at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + or CD33 + /CD38 + /CD123 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment were set to baseline. A – D Concentration-dependent lysis of blasts from AML ( A , B , n = 10) or MDS ( C , D , n = 5) patients by AFM28 compared to RSV/CD16A. E – H Concentration-dependent lysis of blasts from AML patients with low ( E , F , n = 5) or high ( G , H , n = 5) CD64 MFI by AFM28 compared to an Fc-enhanced anti-CD123 IgG antibody. Data in ( A , C , E and G ) are represented as mean ± SD. Data in ( B , D , F and H ) were analyzed using two-way ANOVA and Šídák’s multiple comparisons test. MFI median fluorescence intensity; SD standard deviation.

Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

Techniques: Derivative Assay, Control, Flow Cytometry, Concentration Assay, Lysis, Fluorescence, Standard Deviation

Journal: Nature Communications

Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

doi: 10.1038/s41467-025-63069-y

Figure Lengend Snippet: BMMC patient-derived AML and MDS samples were treated with or without 100 pM of AFM28 for 24 h in the presence of IL-2-preincubated allogeneic NK cells, all derived from different donors, at an E:T ratio of 1:1. Analysis was performed using flow cytometry. LSPCs were defined as viable/CD45 low /CD34 + /CD38 − /CD117 + cells. The gating strategy is shown in Supplementary Fig. . Cell counts of 0 pM treatment for LSPCs and CD123 + LSPCs were set to baseline. A Representative dot plot of LSPC lysis following treatment with 100 pM AFM28 or without AFM28 (indicated as 0 pM) of one AML patient sample. B , C Lysis of LSPCs from AML patients ( B , n = 5) and from MDS patients ( C , n = 5) in the presence of 100 pM AFM28 or without AFM28. Data are represented as mean ± SD and were analyzed using one-way and two-way ANOVA and Šídák’s multiple comparisons test. D , Schematic workflow of the CFU assay. Created in BioRender ( https://BioRender.com/8l2brjy ). E – G CFU assay results of AML ( E , n = 5), MDS ( F , n = 5) and healthy ( G , n = 5) CD34 + cell samples treated with 0/10/100/1000 pM of AFM28 for 24 h in the presence of allogeneic NK cells at an E:T ratio of 1:1. “CD34 + only” describes culturing untreated CD34 + cells without allogeneic NK cells. Colonies were counted manually. Colony count of “CD34 + only” condition was normalized to 100%. Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. SD standard deviation.

Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

Techniques: Derivative Assay, Flow Cytometry, Lysis, Colony-forming Unit Assay, Standard Deviation

Journal: Nature Communications

Article Title: The bispecific innate cell engager AFM28 eliminates CD123 + leukemic stem and progenitor cells in AML and MDS

doi: 10.1038/s41467-025-63069-y

Figure Lengend Snippet: A Fresh whole blood samples from newly diagnosed AML patients ( n = 6) were treated with AFM28 (0/0.01/1 µg/ml equivalent to 0/50/5000 pM) or vehicle in the absence (circles) or presence of healthy donor-derived allogeneic NK cells (squares: fresh, non-expanded NK cells; triangles: cryopreserved cytokine-expanded NK cells generated by a standardized protocol, see Supplementary Methods and Supplementary Fig. ), all derived from different donors, for 24 h at an E:T ratio (NK cells:peripheral leukocytes) of 1:1. Blasts were defined as viable/CD45 low /CD123 + /CD33 + /CD117 + or CD34 + /CD117 + . The gating strategy is shown in Supplementary Fig. . Leukemic blast counts of vehicle treatment without NK cells were set to baseline. E:T ratios of patient’s endogenous NK cells to blasts (in the absence of healthy donor-derived allogeneic NK cells) were 0.1:1, 0.01:1 and 0.2:1 for non-responders and 0.5:1, 0.1:1 and 0.1:1 for responders (data points from top to bottom for 5000 pM AFM28). Data are represented as mean ± SD and were analyzed using one-way ANOVA and Tukey’s multiple comparisons test. B BMMC samples derived from a single AML patient were treated with 0–500 pM of AFM28 for 24 h in the presence of allogeneic AML patient-derived NK cells ( n = 5) or healthy donor-derived NK cells ( n = 5) at a 1:1 E:T ratio. Analysis was performed using flow cytometry. Blasts were defined as viable/CD45 low /CD34 + /CD38 + /CD123 + . Blast counts of 0 pM treatment were set to baseline. HY healthy, SD standard deviation.

Article Snippet: Enrichment of CD34-positive (CD34 + ) cells from BMMCs was performed using the CD34 MicroBead kit (Miltenyi Biotec, cat no. 130-046-702), typically demonstrating > 90% CD34 + cells (data not shown).

Techniques: Derivative Assay, Generated, Flow Cytometry, Standard Deviation