Culturing EBV Producer B95-8 Cells

EBV producer cell line (B95-8 cells) was cultured in RPMI media (GIBCO, Waltham, MA, USA), supplemented with 10% FBS (GIBCO, Waltham, MA, USA), 1% antibiotic and antimycotic solution (Santa Cruz, CA, USA), 50 µg/mL gentamycin (Hyclone, Logan, GA, USA), and 1× glutamine (GIBCO, Waltham, MA, USA) at 37 °C, and 5% CO₂, as previously described [27 (link)]. Cells were cultured until they attained high density (6.5 × 10⁶ cells/mL). Culture supernatants were centrifuged to remove cells/cell debris and then filtered using a 0.2 μm filter. Fresh-filtered virus preparations were used for inoculating rabbits. EBV copy number in the inoculum was estimated by quantitative real-time PCR (qPCR) [27 (link)].