A use of a stem cell conditioned medium to induce ZO-1 proteins expression for skin regeneration, repair and finning is revealed herein. First, mesenchymal stem cells are cultured in a cell culture dish containing complete growth media, wherein the complete growth media include α-MEM, fetal bovine serum, and human-basic fibroblast growth factors. After mesenchymal stem cells are sub-cultured in the complete growth media for three times, a conditioned medium which can effectively increase the activation of tight junction protein Zonula occludens-1 (ZO-1) can be acquired from the basal medium.