Cell Culture Protocols for Various Cell Lines

The human cell lines U937 (myelomonocytic cells, ATCC CRL-1593.2, Manassas, VA, USA) and Jurkat (T cells, ATCC Clone E6-1, TIB-152, Manassas, VA, USA) were cultured under standard cell culture conditions as described previously [38 (link)], In summary, cells were kept in a standard cell culture incubator (5% CO₂, 100% humidity, 37 °C), subcultured every second day and reseeded to a density of 0.2 × 10⁶ cells/mL. For quantification and assessment of cell viability cells were stained with trypan blue. Viability was between 97 and 100%. Population doubling time was between 24 and 28 h. MDA-MB-468 breast adenocarcinoma cells (ATCC HTB-13, Manassas, VA, USA) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing high glucose levels (4.5 g/L) (Gibco/Life Technologies, Hessen, Germany) and supplemented with 10% Fetal Bovine Serum (10270-106, Gibco/Life Technologies, Hessen, Germany) and 1% penicillin/streptomycin (Gibco/Life Technologies, Hessen, Germany). Cells were cultured of a density of 60–70% and medium exchange was performed every 48 h. The viability of all cells in culture was frequently checked.