Cell Culture Protocols for Diverse Cell Lines

HEK293T (CRL-3216), HeLa (CCL-2), HEPG2 (HB-8065) and primary fibroblast cells (CCL-186) were all obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) through LGC Standards (Molsheim, France). The HEK293T stoplight cell line was constructed as described previously [34 (link)]. HEK293T stoplight cells, HEK293T HDR stoplight cells, HEK293T and primary fibroblast cells were cultured in a high-glucose DMEM medium, and HeLa and HEPG2 cells were cultured in Eagle’s minimum essential medium. The culture medium was supplemented with 10% (v/v) FBS (Sigma Zwijndrecht, The Netherlands), and cells were cultured at 37 °C and 5% CO₂. Cell culture plastics, unless otherwise specified, were purchased from Greiner Bio-One (Alphen aan de Rijn, The Netherlands).