Cell Culture Conditions for Virus Research

Porcine embryo kidney (PS) cells were grown in RPMI 1640 (Gibco) supplemented with 5% heat-inactivated fetal calf serum (FCS) at 37°C in a CO₂ (5%) incubator. A549 cells were maintained in Dulbecco’s modified Eagle medium (DMEM) (Life Technologies) supplemented with 10% heat-inactivated FBS (Gibco). Unless otherwise stated, replicon transfected cells were maintained in media supplemented with 2% FCS and 1% Penicillin-Streptomycin (Invitrogen). Sf9 cells (ATCC) were cultured in BioWhittaker Insect-Xpress supplemented with 2% FCS, 1% Penicillin-Streptomycin, and 2.5 μg/mL amphotericin B. Cells were grown at 28°C as monolayers or in suspension with agitation at 100 rpm. Transfection of TBEV replicons were performed in Sf9 monolayer cultures. IRE/CTVM19 was maintained in ambient air at 28°C in L-15 (Leibovitz) medium supplemented with 10% tryptose phosphate broth (TPB), 20% FCS, 2 mM l-glutamine, and Penicillin-Streptomycin (55 (link)).