Quantitative flow cytometric analyses and FACS of human live MNCs were performed using a previously described gating strategy and antigen panel1 (link),15 (link) and antibodies against CD2 (RPA-2.10; 1:20), CD3 (SK7; 1:10), CD14 (MφP9; 1:20), CD19 (SJ25C1; 1:10), CD20 (2H7; 1:10), CD34 (581; 1:20), CD56 (B159; 1:40), CD123 (9F5; 1:20), and CD235a (HIR2; 1:40; all from BD Biosciences, Franklin Lakes, NJ); CD4 (S3.5; 1:20), CD11b (ICRF44; 1:20), CD33 (P67.6; 1:20), and CD90 (5E10; 1:10; all from Thermo Fisher Scientific, Waltham, MA); CD7 (6B7; 1:20) and CD38 (HIT2; 1:20; both from BioLegend, San Diego, CA); CD10 (SJ5-1B4; 1:20; Leinco Technologies, St. Louis, MO); and CD45RA (HI100; 1:10; Tonbo Biosciences, San Diego, CA).
FACS-purified samples were acquired with a BD Influx Cell Sorter (BD Biosciences), and the cell populations were analyzed using FlowJo software (version 10.7.1, Ashland, OR). All experiments included single-stained controls and were performed at MD Anderson’s South Campus Flow Cytometry and Cellular Imaging Facility.
FACS-purified samples were acquired with a BD Influx Cell Sorter (BD Biosciences), and the cell populations were analyzed using FlowJo software (version 10.7.1, Ashland, OR). All experiments included single-stained controls and were performed at MD Anderson’s South Campus Flow Cytometry and Cellular Imaging Facility.