An animal cell culture medium, a preparing method thereof and applications of the culture medium are provided. The culture medium comprises 150-230 parts by weight of amino acids or salts thereof, 180-280 parts by weight of carbohydrates, 160-260 parts by weight of inorganic salts, 1-2 parts by weight of vitamins and 0.002-0.003 part by weight of trace elements. The trace elements comprise manganese chloride tetrahydrate, sodium metavanadate, selenious acid, germanium dioxide, potassium bromide, chromium chloride hexahydrate, ammonium metavanadate, rubidium chloride, cadmium chloride, cobalt chloride hexahydrate, barium acetate, zirconyl chloride octahydrate, sodium fluoride, ammonium molybdate tetrahydrate, copper chloride dihydrate, lithium chloride and aluminium chloride hexahydrate. The culture medium is simple and definite in components, convenient to prepare and use, stable in quality, and low in batch differences, and can be particularly used for culturing a plurality of animal cells. Cell growth states are good and stable.