Three batches of RM, IR-P1 (Merial, France) and BHK-21 were cultured in 48 well plates (Nunc MicroWell Thermo Fisher). RM cells were cultured in RM media; Minimum Essential Medium ((MEM), Thermo Fisher) 5% new born calf serum (Gibco), 20U/mL penicillin (Gibco), 2.5% lactalbumin hydrolysate (Gibco) and sodium bicarbonate (Sigma Aldrich) at pH 7.4 to 7.6) and a seeding density of 1.5x105 cells/mL. IR-P1 cells were propagated in IR-P1 media; MEM (Thermo Fischer), 10% new born calf serum (Gibco), 20U/mL penicillin (Gibco), 2.5% lactalbumin hydrolysate; (Gibco) and sodium bicarbonate (Sigma Aldrich) at pH 7.4 to 7.6) and a seeding density of 2x105 cells/mL. BHK-21 cells were propagated in complete growth medium for BHK cells; alpha MEM (Thermo Fisher) 2 mM-glutamine (Gibco), 20U/mL penicillin (Gibco) and 5% fetal bovine serum (Gibco), at pH 7.2 to 7.4 and a seeding density of 7.5x 105 cells/mL.
Each well of the 48 well plates was inoculated with 200 μL of the cell suspension. The plates were then incubated at 37 °C in a 5% CO2 incubator (Thermo Scientific. Jouan). The plates were monitored daily under the microscope for confluence (Olympus, Model: CKX 31). Cells were infected at 90–100% confluence.
Each well of the 48 well plates was inoculated with 200 μL of the cell suspension. The plates were then incubated at 37 °C in a 5% CO2 incubator (Thermo Scientific. Jouan). The plates were monitored daily under the microscope for confluence (Olympus, Model: CKX 31). Cells were infected at 90–100% confluence.