The invention relates to the technical field of cosmetics and biological medicines, in particular to anti-wrinkle essence containing adipose-derived stem cell extract and a preparation method of the anti-wrinkle essence. The anti-wrinkle essence containing the adipose-derived stem cell extract is composed of the following components in parts by weight: 8 to 20 parts of an adipose-derived stem cellexosome suspension, 8 to 20 parts of an adipose-derived stem cell inclusion, 0.5 to 2 parts of mung bean peptide, 0.5 to 2 parts of tilapia mossambica polypeptide, 0.5 to 2 parts of scorpion venom polypeptide, 1 to 6 parts of glycerinum, 0.1 to 4 parts of ethylene glycol, 0.1 to 4 parts of xanthan gum, 0.1 to 4 parts of hyaluronic acid, 1 to 2 parts of phenoxyethanol, 0.03 to 0.05 part of ethylhexyl, 0.1 to 0.2 part of EDTA, 0.3 to 0.8 part of polysorbate, 1 to 2 parts of oxidized lecithin, 1 to 5 parts of tocopherol and 55.2 to 94.92 parts of distilled water. The anti-wrinkle essence provided by the invention contains the adipose-derived stem cell extract, and a utility evaluation experiment proves that the anti-wrinkle essence has a certain practical effect on skin wrinkle resistance. After the anti-wrinkle essence is used, the improvement conditions of skin elasticity and wrinkle depth of volunteers are superior to those of comparative examples and commercially available anti-wrinkle products, and the anti-wrinkle essence has an obvious synergistic anti-wrinkle effect.