Molm 13 cell line

Cell lines HL-60 and U937 were obtained from American Type Culture Collection (Rockville, MD, USA). MOLM-13 cell line was obtained from Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). HL-60, U937 and MOLM-13 cell lines were cultured in RPMI-1640 medium supplemented with L-glutamine, 1% Penicillin-Streptomycin and 10% (v/v) fetal bovine serum in a humidified 5% CO₂ incubator at 37 °C. Decitabine solution was freshly prepared for each experiment by dissolving decitabine powder in PBS (1X, pH 7.4) at a concentration of 1.00 mM.
For each 1 × 10⁶ cells of HL-60 and U937 cell lines, the dosages of decitabine added to the culture media are 0.00, 0.100, 0.500, 1.00, or 10.0 μM; and were treated for 24 h and 48 h at 37 °C in a humidified 5% CO₂ incubator. MOLM-13 cells (1 × 10⁶ cells) was treated with 0.500 μM of decitabine or an equal volume of PBS (1X, pH 7.4) as control for 0, 4, 8, and 24 h at 37 °C in a humidified 5% CO₂ incubator. After treatment, the cells were removed from the medium by centrifugation at 1,500 × g, 4 °C for 5 min; and washed twice with 5.00 mL of PBS (1X, pH 7.4) each. The cell pellets were collected and stored in −20 °C until DNA extraction.