The invention discloses a non-animal-derived culture medium, prepared from the following components: L-arginine, L-asparagine, L-aspartic acid, L-cystine dihydrochloride, L-glutamate, glycine, L-histidine, L-hydroxyproline, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-glutamine, succinic acid, sodium pyruvate, p-aminobenzoic acid, calcium nitrate, potassium nitrate, ammonium nitrate, anhydrous magnesium sulfate, anhydrous sodium dihydrogen phosphate, sodium hydrogen carbonate, potassium chloride, sodium chloride, magnesium chloride, anhydrous calcium chloride, glucose, ribose, reduced glutathione, phenol red, biotin, D-calcium pantothenate, folic acid and nicotinamide. The non-animal-derived culture medium can be used for culturing NK cells, T cells, mesenchymal stem cell series, CHO cell series, Vero cell series, 293 cell series, K562 cell series, human skin keratinocyte series and fibrous blastocyte series.