The invention discloses an umbilical cord mesenchymal stem cell preparation and a preparation method and application thereof. Through exploration and verification by a large quantity of experiments, the 3rd-10th-generation umbilical cord mesenchymal stem cells are selected as raw materials, and by a method of dispersing tissue by a mechanical method and then performing treatment with mixed enzymes containing collagenase I, collagenase II and trypsin for a short time for many times, the vitality of the separated umbilical cord mesenchymal stem cells is guaranteed. A culture medium special for the umbilical cord mesenchymal stem cell preparation, containing DMEM/F12 (without phenol red), 10% FBS, 20 kinds of amino acids and 8 kinds of vitamins, is adopted, mixed liquor of repeated cell culture supernatant and cell lysate supernatant is collected, more umbilical cord mesenchymal stem cell active protein and active factors are obtained, the preparation cost is reduced, and industrial requirements for a stem cell product preparation are met. The prepared umbilical cord mesenchymal stem cell preparation can promote proliferation of epidermic cells and skin fibroblast, is safe and is free from toxic and side effects, an allergy does not exist, and when the umbilical cord mesenchymal stem cell preparation is applied to feature-beautifying skin-care products, the effects are notable.