Phosphate buffered saline (pbs)

FCM analysis was performed as we recently reported [20 (link), 22 (link)]. In brief, the fresh-enriched MNCs (day 0) and the MNC-derived cells (day 14) were harvested by centrifugation at 300×g for 5 min and resuspended by 1 × PBS (Solarbio, China) for twice. After that, the cells were incubated in 1 × PBS (Solarbio, China) with 2% fetal bovine serum (FBS) (Australia) and the fluorescence-conjugated antibodies such as anti-CD3-PE (BioLegend, USA), anti-CD3-APC (BioLegend, USA), anti-CD4-PE (BioLegend, USA), anti-CD8-PE-Cy7 (BioLegend, USA), anti-CD56-APC (BioLegend, USA), anti-CD16-FITC (BioLegend, USA), anti-CD25-FITC (BioLegend, USA), anti-NKG2D-perCP-Cy5.5 (BioLegend, USA), anti-NKp44-APC-Cy7, anti-NKp46-PE-Cy7 (BD Biosci, USA), anti-NKG2A-PE (BD Biosci, USA), anti-CD107a-PE (BD Biosci, USA), 7-AAD (BD Pharmigen), Propidium iodide (PI) (BD Pharmigen, USA) or Annexin V-FITC (Tianjin Sungene Biotech, China) in dark for 30 min. Finally, the cells were washed and turned to FACS Canto II (BD Biosci, USA) and FlowJo 10.0 software (Tree Star, USA) for analysis. The list of the indicated antibodies was available in Additional file 1: Table S3.

Cell cycle assessment of the MNCs (HD-MNCs, AML-MNCs) and the derivatives at the indicated time points was conducted as we reported [19 (link), 23 (link)]. In brief, the cells were harvested and suspended in 1 × PBS (Solarbio, China), and then incubated with anti-CD3-PE and anti-CD56-APC antibodies for 20 min in dark. After that, the cells were incubated in 70% (v/v) ethanol (Thermo Fisher Scientific, USA) and fixed for 24 h, and followed by centrifugation at 1000×g for 5 min and resuspended with 1 × PBS (Solarbio, China) at 4 ℃ for twice. Then, the cells were incubated with PI staining solution (BD Pharmigen, USA) for 30 min at 37 ℃ and detected by BD LSR II (BD Biosci, USA) and the ModFit software (Verity Software House Co. Ltd, USA).