FIELD : medicine; biotechnology.SUBSTANCE : invention relates to medicine and biotechnology and is intended for the cultivation of animal cells in vitro in the production of virus vaccines. Method of decontamination of nutrient media for cultivation of animal cells in vitro comprises heat treatment of the nutrient medium before irradiation by heating to a temperature of 55–60?°C for 25–30 minutes, and the irradiation is carried out in a dose (0.8–1.5)×103 Gy by gamma rays.EFFECT : use of the invention provides the increased efficiency of the decontamination of nutrient media for cultivation of animal cells in vitro.1 cl, 4 ex