Working with mammalian cell lines

SP Sebastian Pilsl
CM Charles Morgan
MC Moujab Choukeife
AM Andreas Möglich
GM Günter Mayer
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HEK293 cells (CLS Cell Lines Service) were cultured in DMEM medium (high glucose, GlutaMAX), supplemented with 1% non-essential amino acids (NEAA), 1% Sodium Pyruvate (Thermo Fisher Scientific) and 10% fetal calf serum (FCS, Sigma-Aldrich), at 37 °C in a humidified 5% CO2 atmosphere and were passaged every 2–3 days. HEK293PAL cells were cultured 1 week before and after cell sorting with 1% penicillin/streptomycin and in presence of Geneticin (G418, 400 µg mL−1, Thermo Fisher Scientific). Mycoplasma testing was performed every three months using PCR detection (Minerva biolabs).

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