Culturing Influenza Virus in MDCK Cells

Madin-Darby canine kidney cells were purchased from the Pasteur institute of Iran, Tehran. The cells were grown at 37 °C in 5% CO₂ with 85% of humidity in DMEM(BIO-IDEA), supplemented with 10% of Fetal Bovine serum (DNABIOTECH) and 1% of penicillin (BIO-IDEA). The influenza virus vaccine strain, A/Puerto Rico/8/1934 (H1N1) (ATCC VR-897™), sourced from the Influenza Department at the Pasteur Institute of Iran, was cultured in MDCK cells. The virus was cultured in MDCK cells with the addition of 1 μg/ml of trypsin-Tosylamide Phenylethyl Chloromethyl Keton-treated Trypsin (TPCK) from Sigma, USA. After a 48-hour incubation period, the supernatant containing the virus was collected and the Reed and Muench formula was used for virus titration [24 (link)].