Primary AML and MDS cells were collected from patients who had consented to research protocols approved by the Institutional Review Board at The University of Texas MD Anderson Cancer Center for analysis of hematologic malignancies. BM aspirate mononuclear cells from patients with MDS were purified by Ficoll density centrifugation using standard procedures. Lymphocytes were isolated by using Lymphocyte Separation Medium (SIGMA).
Primary AML peripheral blood mononuclear cells (PBMCs) were cultured in serum-free Expansion Medium (Cat. 09650) supplemented with BIT 9500 Serum Substitute (Cat. 09500; both from STEMCELL Technologies Inc., Vancouver, BC, Canada) and cytokines including stem cell factor (SCF, 100 ng/mL, Cat. 300-07), Flt3 ligand (50 ng/mL, Cat. 300 – 19), IL3 (20 ng/mL, Cat. 200-03), and G-CSF (20 ng/mL, Cat. 300 – 23; all from Peprotech, Rocky Hill, NJ) and StemRegenin 1 (1 μM, Cat. S2858; Selleck Chemicals LLC, Houston, TX).
Primary AML peripheral blood mononuclear cells (PBMCs) were cultured in serum-free Expansion Medium (Cat. 09650) supplemented with BIT 9500 Serum Substitute (Cat. 09500; both from STEMCELL Technologies Inc., Vancouver, BC, Canada) and cytokines including stem cell factor (SCF, 100 ng/mL, Cat. 300-07), Flt3 ligand (50 ng/mL, Cat. 300 – 19), IL3 (20 ng/mL, Cat. 200-03), and G-CSF (20 ng/mL, Cat. 300 – 23; all from Peprotech, Rocky Hill, NJ) and StemRegenin 1 (1 μM, Cat. S2858; Selleck Chemicals LLC, Houston, TX).
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