The present study was designed in a trial to use serum replacement instead of the newborn calf serum in propagation of BHK-21 cell cultures with subsequent reducing the cost of foot and mouth disease (FMD) vaccine production. Two batches of BHK-21 cell culture were prepared where the medium of the first batch was supplemented with newborn calf serum while the medium of the second batch was supplemented with serum replacement. FMD virus was propagated 7 passages using BHK-21 cell culture. Both virus titration and complement fixation titer (CF) revealed that propagation of FMD virus in cell cultures supplemented with newborn calf serum yields a titre higher than that in case of cells supplemented with serum replacement. Also two batches of FMD inactivated vaccine were prepared from the virus propagated in the two-mentioned cell culture batches. Two groups of susceptible calves were vaccinated with these vaccines. Both of virus neutralization test (VNT) and enzyme linked immunosorbent assay (ELISA) revealed that higher antibody levels were induced in calves vaccinated with the vaccine prepared from cells supplemented with calf serum than those vaccinated with vaccine prepared from cells grown with serum replacement. BHK-21 cell culture supplement with newborn calf serum is most susceptible for FMD virus propagation yielding higher titer of the virus. Moreover, the growth pattern of the used cell culture was much better when the newborn calf serum was used.