The invention discloses a preparation method and application of stem cell culture solution containing wash-free mask liquid. The mask liquid is prepared from, by weight, 0.2%-0.5% of high molecular weight sodium hyaluronate, 1%-5% of glycerin, 0.1%-2.0% of a stem cell culture solution, 0.05%-0.5% of nanosized sodium hyaluronate, 0.3%-1% of 1,2-hexanediol, 0.3%-0.5% of p-hydroxyacetophenone, 0.05%-0.2% of xanthan gum, 1%-5% of moringa seed extracts and the balance deionized water. Accordingly, the mask liquid contains the stem cell culture solution which contains a larger number of growth factors, ultra-low molecular weight nanosized sodium hyaluronate and the moringa seed extracts can make the skin care effect of the stem cell culture solution brought into full play, stem cell growth factors go deep into the skin dermis and subcutaneous tissue to comprehensively activate stem cell differentiation inside the skin when nanosized sodium hyaluronate is subjected to transdermal absorption, stem cell differentiation inside the skin is comprehensively activated, senescent cells are rapidly updated and replaced, the skin is improved, moisturizing and elasticity are improved, and wrinkles are reduced. Accordingly, cleaning is not needed after a mask is used.