Propagation of HSV-1 and Bovine Coronavirus in Cell Cultures

HSV-1 McKrae [27 (link)] and HSV-1-GFP [28 (link)] were cultured in Vero cells in Eagle’s Minimum Essential Medium (EMEM) supplemented with 5% fetal bovine serum (FBS) (GeminiBio, Sacramento, CA, USA), penicillin (100 U/mL), and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc., St. Louis, MO, USA). Vero cells were cultured in EMEM supplemented with 10% fetal bovine serum (FBS) (GeminiBio, USA), penicillin (100 U/mL), and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc., St. Louis, MO, USA) at 37 °C with 5% CO₂ in a humidified incubator as we described previously [22 (link),29 (link)].
Bovine coronavirus (BCoV) was the NADL-Nebraska Strain isolated by the National Veterinary Services Laboratories (NVSL) in 1981 [30 (link)]. BCoV was grown on human rectal tumor (Hrt-18G) cells [30 (link),31 (link)], which were maintained in EMEM supplemented with 10% FBS (GeminiBio, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc.) at 37 °C with 5% CO₂ in a humidified incubator. The virus was propagated in EMEM supplemented with 2.5 μg/mL trypsin and 2.5 μg/mL pancreatin, 1× insulin–transferrin–selenium (Cat No. 51500-056, GIBCO) [31 (link)].