The invention belongs to the technical field of cell culture media, and particularly relates to an amniotic cell culture medium and a preparation method thereof. The amniotic cell culture medium provided by the invention comprises a basal culture medium and additives, wherein the additives are added into the basal culture medium, and the additives include the components according to final concentration: 10 to 15 mum of linoleic acid, 15 to 18 mug/mL of beta-carotene, 3 to 6 mum of palmitic acid, 3 to 5 mum of L-glutamine, 3 to 7 mg/mL of polylactic acid-glycolic acid copolymer, 0.001 to 0.003 mug/mL of beta-mercaptoethanol, 2 to 5 mg/mL of human albumin, 22 to 27 mug/mL of vitamin B, 18 to 25 ng/mL of epidermal growth factor and 0.7 to 1.1 mug/mL of insulin. The amniotic cell culture medium does not contain serum, and the amniotic cell clone number and cell proliferation rate in vitro proliferation can be quickly and effectively increased.