Abstract: Defined serum-free, low protein media (LPKM), that supports 1) Vero cell growth for up to 20 passages, 2) Vero cell growth on microcarriers and 3) rotavirus production is provided. Maximum cell densities attained are 60-100% of that in serum-containing medium; the doubling time is equal to that for cells in serum containing medium. Rotavirus titers achieved in LPKM-1 are 50-100% of the serum-containing process. Finally, since LPKM-1 contains no animal-sourced proteins, the problems associated with the serum-containing rotavirus production process (i.e. lengthy wash steps before infection, potential introduction of adventitious agents and lot-to-lot variability) can be avoided; while maintaining nearly equivalent product titers.