Cell Culture Protocols for Diverse Cell Lines

Human HAP1 cells were obtained from Horizon Discovery, they were grown in Iscove’s Modified Dulbecco’s Medium (IMDM) from GIBCO^®, containing l-Glutamine and 25 mM HEPES and supplemented with 10% fetal bovine serum (FBS) and 1% Penicillin/Streptomycin (P/S). HEK293T cells were obtained from the CRUK Cell Facility, they were used for virus production, expanded in Dulbecco’s modified Eagle medium (DMEM) and supplemented with 10% FBS. U2OS cells were originally obtained from ATCC cell repository, they were cultured in DMEM (Sigma-Aldrich), supplemented with 10% FBS. Human mCherry-Geminin-expressing U2OS^{44 (link)}, FANCC-deficient VU1131 and complemented VU1131 cells (a gift from Josephine Dorsman, VU Medical Center Amsterdam) were cultured in DMEM, supplemented with antibiotics and 10% fetal calf serum. VU1131 cells were grown with G418 (300 μg/mL). All cells were grown at 37 °C in a 3% oxygen and 5% CO₂ atmosphere. All cell lines used in this publication were tested negative for mycoplasma contamination using the MycoAlert™ Mycoplasma Detection Kit. They were all authenticated by the specified providers and, furthermore, they are not listed as commonly misidentified by ICLAC.