Flow cytometry analyses on patients’ samples
CD41/CD61-PC7, CD19-APCR700
Identification and quantification of HSPC subpopulations from BM-GT and Pt1 were previously performedMouse anti-human CD3-BV605 (Clone: OKT3; Biolegend, 317322), Verified Reactivity: Human; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:50Mouse anti-human CD56-PC5 (Clone: 5.1H11; Biolegend, 362516), Verified Reactivity: Human; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:50Mouse anti-human CD14-BV510 (Clone: M5E2; Biolegend, 301842), Verified Reactivity: Human, Cynomolgus, Rhesus; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:50Mouse anti-human CD33-BB515 (Clone: WM53; BD Biosciences, 564588), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:50Mouse anti-human CD41/CD61-PC7 (Clone: A2A9/6; Biolegend, 359812), Verified Reactivity: Human; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:50Mouse anti-human CD66b-BB515 (Clone: G10F5; BD Biosciences, 564679), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:50Mouse anti-human CD7-BB700 (Clone: M-T701; BD Biosciences, 566488), Verified Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Reported); Application: Flow cytometry (Routinely Tested). Dilution: 1:50Mouse anti-human CD45-BUV395 (Clone: HI30; BD Biosciences, 563792), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:33Mouse anti-human CD38-BUV737 (Clone: HB7; BD Biosciences, 612824), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:33Mouse anti-human CD90-APC (Clone: 5E10; BD Biosciences, 559869), Verified Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon, Pig, Dog (Tested in Development); Application: Flow cytometry (Routinely Tested). Dilution: 1:33Mouse anti-human CD135-PE (Clone: BV10A4H2; Biolegend, 313306), Verified Reactivity: Human; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:33Mouse anti-human CD11c-BV650 (Clone: B-ly6; BD Biosciences, 563404), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:20Mouse anti-human CD10-BV786 (Clone: HI10a; BD Biosciences, 564960), Verified Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development); Application: Flow cytometry (Routinely Tested). Dilution: 1:20Mouse anti-human CD34-BV421 (Clone: 561; Biolegend, 343610), Verified Reactivity: Human; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:20Mouse anti-human CD45RA-APCH7 (Clone: HI100; Biolegend, 304128), Verified Reactivity: Human; Reported Reactivity: Chimpanzee; Application: Flow cytometric analysis of antibody surface-stained cells. Dilution: 1:20Mouse anti-human CD71-BV711 (Clone: M-A712; BD Biosciences, 563767), Verified Reactivity: Human (QC Testing); Application: Flow cytometry (Routinely Tested). Dilution: 1:20Mouse anti-human CD19-APCR700 (Clone: SJ25C1; BD Biosciences, 659121), Verified Reactivity: Human; Application: Flow cytometry. Dilution: 1:20All the antibodies were purchased from Biolegend and BD Biosciences and they are well-characterized and validated by providers. Titration assays were performed to assess the best antibody concentration. After surface marking, the cells were incubated with PI (Biolegend) to stain dead cells. Absolute cell quantification was performed by adding Flowcount beads (BD Biosciences) to samples before WBD procedure. All stained samples were acquired through BD LSR-Fortessa or BD Symphony A5 (BD Biosciences) cytofluorimeters after Rainbow beads (Spherotech) calibration. Raw data were collected through DIVA software Version 8.0.2 and analyzed with FlowJo software Version 10.5.3 (BD Biosciences). Technically validated results were always included in the analyses, and we did not apply any exclusion criteria for outliers.