The invention provides a preparation of stem cell active factor freeze-dried powder combined with traditional Chinese medicine for the treatment of mild to moderate acne and a preparation method of the preparation, and relates to the field of stem cell preparations and application of traditional Chinese medicine. The problem of poor effect of existing preparations for acne treatment is solved. Thepreparation is prepared from the stem cell active factor freeze-dried powder, a honeysuckle extracting solution and mixed traditional Chinese medicine powder; the mixed traditional Chinese medicine powder is prepared from scutellaria baicalensis georgi, angelica dahurica, rhizoma typhonii, radix sileris, salviae miltiorrhizae and mung beans. The preparation method includes the steps that umbilical cord and adipose derived mesenchymal stem cells are taken and mixed and cultured, and supernatants of P3, P4, P5 and P6 cells are obtained; the supernatants of the P3, P4, P5 and P6 cells are mixedand filtered, mannitol is added, sub-packaging and precooling are performed, and the freeze-dried powder is prepared; the honeysuckle extracting solution is prepared; and the mixed traditional Chinesemedicine powder is prepared. The preparation has a good effect on treating the mild to moderate acne, is not easy to produce dependence, acne marks can be eliminated or the production of acne and whelk can be inhibited, and the occurrence of fine wrinkles can be reduced and inhibited. The preparation method is simple and easy to operate, low in cost, and has non-toxic side effects. The preparation prepared by the method is used for treating the mild to moderate acne.