Culturing HCT116 and HEK293T Cell Lines

HCT116 colorectal human cells (American Type Culture Collection—ATCC, Manassas, VA) which are near diploid with 45 median chromosome count were cultured at 37 °C and 5% CO₂ in complete DMEM medium (DMEM with 4.5 g/L glucose (Gibco, Waltham, MA) supplemented with 10% (v/v) fetal calf serum (Eurobio, Les Ullis, France), 1% (vol./vol.) GlutaMAX^® (Gibco) and 1% (v/v) penicillin/streptomycin (Gibco)). A subculture of HEK293T human embryonic kidney cells was derived at Genethon and used to produce LV for research or for clinical use as previously reported [4 (link)]. Such cells were derived from a working cell bank which was authenticated and tested negative for mycoplasma.