Cell Culture Maintenance Across Species

Canine MDCKII^{27 (link)}, porcine NSK and NPTr cells^{31 (link)} and human A549^{32 (link)} and HEK293T cells^{33 (link)} were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 units per mL of penicillin and 100 μg per mL of streptomycin. Chicken hepatocellular epithelial cell line (LMH) ^{34 (link)} and chicken fibrolast cells (DF-1) ^{35 (link)} were grown in DMEM supplemented with 8% (v/v) fetal calf serum, 2% (v/v) chicken serum, 2 mM L-glutamine, 100 units per mL of penicillin and 100 μg per mL of streptomycin. Bat cells EpoNi/22.1^{36 (link)} and Carper AEC B-3^{37 (link)} were maintained in DMEM supplemented with 10% (v/v) fetal calf serum (FCS), 2 mM L-glutamine, 100 units per mL of penicillin, 100 μg per mL of streptomycin, 1 mM sodium pyruvate and 0.1 mM non-essential amino acids (NEAA, # NEAA-10201–100). Primary human bronchial airway epithelial cells (MucilAir™, Epithelix Sàrl) reconstituted in Transwells of 6.5 mm of diameter with a pore size of 0.4 μm (Costar 3470, Corning incorporated) were maintained in MucilAir™ Culture Medium.