Evaluating Combination Drug Treatments on AML Cell Lines

Both cell lines were cultured in vitro to assess drug treatment, by plating the OCI/AML3 (DSMZ–Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH–German Collection of Microorganisms and Cell Cultures) and THP1 (ATCC–American Type Culture Collection) cells in 96‐well plates, at 10⁴ cells/200 µL/well in 2 types of media: 80% alpha‐MEM (Invitrogen) with 20% foetal bovine serum (FBS) (Invitrogen) for OCI/AML3 or RPMI1640 (Invitrogen) with 10% FBS, 2 mM L‐glutamine for THP1, and then treating them for 48 hour with 37.5 µM olaparib (Selleckchem), 100 µM cytarabine (Sigma‐Aldrich), 1.4 µM daunorubicin (Sigma‐Aldrich) and 10 µM azacytidine (Sigma‐Aldrich), either alone or in combination. Cells were cultured in an incubator at 37°C and 5% CO₂, as previously described.²⁴ (link), ²⁵ (link), ²⁶ (link) Cell proliferation was evaluated by using the CellTiter 96® AQueous Non‐Radioactive Cell Proliferation Assay (Promega) and analysed with BioTek Synergy H1 Hybrid Multi‐Mode Reader (BioTek Instruments). All reagents and compounds were purchased from Invitrogen and had a 99.9% purity. The in vitro experiments were carried out after the approval of the Ethics Committee of the Iuliu Hatieganu University of Medicine and Pharmacy in Cluj Napoca.