Cell Culture Protocols for KSHV-Infection

L1T2 cells, obtained from ATCC, were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin and passaged every 3–4 days. At the onset of this project, L1T2 cells were available for purchase from ATCC but are no longer available despite having been demonstrated to maintain long-term KSHV infection and to induce tumors upon xenotransplantation into immune deficient mice [20 (link)]. TIVE cells, kindly provided by Rolf Renne, were cultured in Medium 199 supplemented with 20% FBS, 1% penicillin/streptomycin, 1% 200 mM L-Glutamine, and 30 mg Endothelial Cell Growth Factor (Sigma, Burlington, MA, USA) and reportedly lack the capacity to produce tumors in immunodeficient mice [19 (link)]. The media were changed two times per week, and the cells were split 1:2 when the culture reached ~60% confluency.