Minimal essential media

C7/10 Aedes albopictus cells were grown at 28 °C under 5% ambient CO₂ in 1× Minimal Essential Media (Corning) supplemented with 10% heat-inactivated fetal bovine serum (Corning), 1% L-glutamine (Corning), 1% non-essential amino acids (Corning), and 1% antibiotic–antimycotic solution (Corning). Vertebrate baby hamster kidney fibroblast or BHK-21 cells, as well as human embryonic kidney epithelial or HEK293 cells, were grown at 37 °C under 5% ambient CO₂ in the same 1× Minimal Essential Media (Corning) as the C7/10 cells.

HSV1 KOS (a kind gift from David Bloom, Univ. of Florida College of Medicine). Vero cells (ATCC) were maintained with Minimal Essential Media (Cellgro) supplemented with 100 IU penicillin/ml, 100 microgram streptomycin/ml, 2.5 microgram amphotericin B/ml, and 10% heat-inactivated fetal bovine serum (Hyclone). Cells were incubated at 37°C, 5% CO2 in a humidified chamber. Plaque reduction assays were performed by diluting virus stocks and preincubating 100–200 plaque-forming units (pfu) with increasing concentrations of botanicla extract for 20 minutes. Monolayers were infected for 1 hour at 37°C followed by incubation in media containing the botanical for 3 days at 37°C. Plaques were visualized by staining with 0.1% crystal violet in 20% ethanol.

Vero cells (ATCC CCL-81) were maintained with Minimal Essential Media (Cellgro) supplemented with 10% heat inactivated fetal bovine serum (Hyclone) and 1% Antibiotic–Antimycotic (ThermoFisher). Cells were incubated at 37 °C, with 5% CO₂ in a humidified chamber. HSV-1 KOS (a kind gift from David Bloom, Univ. of Florida College of Medicine) was propagated in Vero cells.