Cell Culture of Haploid Leukemia and Endothelial Cells

Myelogenous leukemia-derived, near-haploid human HAP1 cells (44 (link)) were purchased from the American Type Culture Collection and cultured in Iscove's modified Dulbecco's medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin (Merck). HUVECs were purchased from Cell Applications and grown in endothelial cell basal medium 2 supplemented with SupplementPack Endothelial Cell GM2 (PromoCell) on collagen-coated dishes. MEDEP-E14 cells (45 (link)) were obtained from RIKEN BioResource Research Center and grown in Iscove's modified Dulbecco's medium containing 15% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 1× ITS Liquid Media Supplement (Merck), 0.45 mM α-monothioglycerol (Merck), 50 mg/ml ascorbic acid (Merck), and 3 units/ml human erythropoietin (Kyowa Kirin). Cells were cultured at 37 °C under 5% CO₂. Transfections were performed using the reagents Lipofectamine and PLUS (Thermo Fisher Scientific), according to the manufacturer's instructions.