Vero cells (ATCC CCL-81) were cultured in alpha minimum essential medium (α-MEM; Invitrogen, Carlsbad, CA) with 5% fetal bovine serum (FBS; Invitrogen) and antibiotic-antimycotic solutions (100×; Invitrogen) and maintained at 37 °C in a humidified 5% CO2 incubator. Virulent Korean PEDV strain KOR/KNU-141112/2014 was isolated and propagated in Vero cells as described previously (Lee et al., 2015 (link)). A viral stock was prepared from the 5th passage in cell culture (KNU-141112-P5) and used as the parental virus in this study (GenBank accession no. KR873434). PEDV N protein-specific monoclonal antibody (MAb) was obtained from ChoogAng Vaccine Laboratory (CAVAC; Daejeon, South Korea).
Antibiotic antimycotic solutions 100
Manufactured by Thermo Fisher Scientific
Sourced in United States
Antibiotic-antimycotic solutions (100×) are a sterile, liquid cell culture reagent used to prevent bacterial and fungal contamination in cell culture applications. The solutions contain a combination of antibiotics and antifungal agents at a 100× concentration, which can be added to cell culture media to inhibit the growth of a broad spectrum of microorganisms.
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2 protocols using antibiotic antimycotic solutions 100
1
Propagation of Virulent PEDV Strain
2
SADS-CoV Infection of Epithelial Cells
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Porcine ileum epithelial (IPI-2I) cells, African green monkey kidney (Vero E6) cells, and human embryonic kidney (HEK293T) cells were cultured in Dulbecco’s minimal essential medium (DMEM; Invitrogen) with 10% fetal bovine serum (Invitrogen, Waltham, MA, USA) and antibiotic–antimycotic solutions (100×; Invitrogen). The cells were maintained at 37 °C in a humidified 5% CO2 incubator. SADS-CoV was maintained in our laboratory. SADS-CoV infected IPI-2I and Vero E6 cells as described previously [26 (link)].
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