Effect of cultured medium of human umbilical cord blood-derived mesenchymal stem cells on melanogenic enzyme activity in mouse B16 melanoma cells

Abstract

The cultured medium of mesenchymal stem cell is a valuable resource for a variety of stem cell-derived cytokines and growth factors with a potential therapeutic effect on skin diseases. Here, we investigated the effect of the cultured medium of human umbilical cord blood-derived mesenchymal stem cell (CM-hCBSC) on melanogenesis in mouse B16 melanoma cells stimulated by α-melanocyte stimulating hormone (α-MSH). Our results show that CM-hCBSC inhibits melanin synthesis due to the decrease in cellular tyrosinase activity in B16 melanoma cells without cytotoxicity. The activity of melanogenic enzymes such as tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2 were evaluated by western blot and semi-quantitative RT-PCR. The CM-hCBSC down-regulated the protein and mRNA expression levels of tyrosinase and TRP-1 in a dose-dependent manner. In addition, TRP-2 protein and mRNA expression levels were found to be down-regulated to a lesser extent. We also measured the level of cytokines in CM-hCBSC using the Luminex total system. It was found that the top enriched cytokine in the CMhCBSC was interukine-6 (IL-6), known as a potent regulator of melanin synthesis together with IL-1α and tumor necrosis factor α (TNF-α). These results indicate that secretary factors including IL-6 in the CM-hCBSC may inhibit melanin synthesis by down-regulating the expression of tyrosinase and TRP1. This study suggests that CMhCBSC has a therapeutic effect as an anti-melanogenic agent and may be effective against hyperpigmentation disorders.