Spodoptera frugiperda sf9 cells

Swine pulmonary alveolar macrophage (CRL2845), human embryonic kidney cells (HEK293), Madin–Darby canine kidney (MDCK), swine testis cells (ST), and African green monkey kidney cells (Vero) were cultured with Dulbecco’s Modified Eagle Medium (DMEM, HyClone, Logan, UT, USA), supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA), and 1% penicillin/streptomycin (Cytiva, Lewisville, TX, USA) at 37 °C with 5% CO₂. Spodoptera frugiperda (Sf9) cells (Invitrogen, Carlsbad, CA, USA) were cultured in SF-900™ II SFM medium (Gibco, Grand Island, NY, USA) supplemented with 10% FBS and incubated at 27 °C.

The 293T and Huh7 cells (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Corning-Costar, Coring, NY, USA) supplemented with 10% fetal bovine serum and penicillin-streptomycin (FBS; Gibco, Grand Island, NY, USA). These cells were maintained at 37 °C. The Spodoptera frugiperda (Sf9) cells (Invitrogen, San Diego, CA, USA) were cultured in suspension in SF-900 II serum-free medium (Invitrogen, San Diego, CA, USA) supplemented with 10% FBS and penicillin-streptomycin. These cells were maintained at 27 °C.
Female BALB/c mice, 6–8 weeks old, were purchased from Changchun Institute of Biological Products (Changchun, China). All experiments involving mice adhered to the principles of the Welfare and Ethics Committee of the Military Veterinary Research Institute at the Academy of Military Sciences. The mice were provided ad libitum access to sterilized water and food throughout the study and were vaccinated under BSL-2 conditions.
Healthy male horses 2–6 years old and 400–500 kg were provided by Red Hill Military horse farm (Changchun, China). Horse studies were conducted with prior approval from the Animal Welfare and Ethics committee of the Institute of Military Veterinary, Academy of Military Medical Sciences (permit number SCXK-2012-017), according to Horse Quarantine and Immunization Protocols for Equine Serum Production.

Spodoptera frugiperda Sf9 cells (Invitrogen) were maintained in Sf-900 III Serum Free Medium (Gibco) and incubated at 27 °C, 140 rpm without CO₂ exchange in a non-humidified orbital shaker. Suspension culture was passaged when reaching a density of 2E6 viable cells/mL and was seeded at 0.5E6 viable cells/mL. Cell counting was performed on the suspension culture using Trypan Blue (Invitrogen) to determine the cell density and viability at every passage.