Characterization of AML Patient Samples

Primary AML patient samples and human umbilical cord blood samples were obtained from the First Hospital of Jilin University. Normal peripheral blood mononuclear cells (PBMCs) were donated by healthy individuals. Informed consent was obtained in all cases as per the Declaration of Helsinki. Both the study and patient sample collection were approved by the Human Ethics Committee of The First Hospital of Jilin University. All AML patient samples were screened for the following gene mutations via PCR amplification and automated DNA sequencing: FLT3-ITD, NPM1, C-kit, CEBPA, IDH1, IDH2, SF3B1, TP53, ZRSR2, GATA2, KMT2A, SH2B3, TCF, TET2, RUNX1, and DNMT3A. Cytogenetics and detection of fusion genes via real-time PCR were also performed, as previously described^{16 (link),17 (link)}. Characteristics of the individual AML patients are listed in Supplementary Table S1. Primary patient samples were purified with Ficoll-Hypaque density centrifugation, and then cultured in RPMI 1640 media plus 20% fetal bovine serum, ITS Solution (Sigma-Aldrich, St Louis, MO, USA), and 20% supernatant of the 5637 bladder cancer cell line (to provide sources of granulocyte–macrophage colony-stimulating factor, granulocyte colony-stimulating factor, interleukin-1 beta, macrophage colony-stimulating factor, and stem cell factor)^{16 (link),18 (link),19 (link)}.