Skm 1

The human AML cell lines MOLM13, THP-1, OCI-AML3, and SKM-1 were obtained from ATCC (American Type Culture Collection) in 2011. The MV4-11 cell line was purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zelkulturen, Braunschweig, Germany) in 2011. All human cell lines were authenticated by short tandem repeat DNA profiling at CCTCC (China Center for Type Culture Collection). The SKM-1 FLT3-ITD knock-in mutant cell lines were generated using the CRISPR/Cas9 system according to the reported method.^{46 (link),47 (link)} All cell lines except MV4-11 and OCI-AML3 were cultured in RPMI 1640 medium (Gibco, Waltham, MA); MV4-11 and OCI-AML3 cells were maintained in Iscove’s modified Dulbecco’s medium (Gibco) and a-Minimum Essential Medium (Gibco), respectively. All media contained 10% fetal bovine serum (Gibco). Cells were maintained in a 37 °C humidified atmosphere containing 5% CO₂.

Human AML cells SKM-1 and THP-1 were obtained from the China Center for Type Culture Collection (Wuhan, China). All cell lines were tested and authenticated, utilizing short tandem repeat profiling every six months. SKM-1 and THP-1 cells were cultured in RPMI-1640 (GIBCO, Life Technologies Corporation, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum and 100 U/mL of penicillin-streptomycin at 37°C and 5% CO₂ and normal O₂. The CCK8 assay (A311-01/02, Vazyme, China) was used to test the proliferation of cells, according to the manufacturer’s instructions.

Human MDS cell lines (SKM-1 and MDS-L) and HEK293T cell lines were purchased from ATCC (Manassas, VA, USA). SKM-1 cells were maintained in RPMI-1640 (Gibco, Rockville, MD, USA). MDS-L cells were cultured in RPMI-1640 medium plus 50 μM 2-mercaptoethanol and 100 U/ml IL-3. HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco). All media were supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin–streptomycin (Gibco). All cells were cultured at 37 °C in a 5% CO₂ atmosphere. The cells were authenticated using STR profiles. All cells were routinely tested as mycoplasma-free. All clinical samples were obtained with informed consent at the First Affiliated Hospital of University of South China (Hengyang, China) and Xiangya Hospital of Central South University (Changsha, China). Sample collection was approved by the Hospital’s Protection of Human Subjects Committee. MDS primary samples were cultured in RPMI-1640 medium containing 20% fetal bovine serum and 100 U/ml IL-3.