Collectively, trypsin-dependent PEDV isolates 2013-A and NJ could effectively replicate in the Vero/TMPRSS2 and Vero/MSPL cells without trypsin, indicating that the Vero/TMPRSS2, especially Vero/MSPL cells, can contribute to the propagation and infection of PEDV isolates without trypsin supplementation. Here, TMPRSS2 is linked to infection.