Because adipose-derived stem cells (ASCs) are usually expanded to acquire large numbers of cells for therapeutic applications, it is important to increase the production yield and regenerative potential during expansion. Therefore, a tremendous need exists for alternative ASC stimuli during cultivation to increase the proliferation and adipogenic differentiation of ASCs. The present study primarily investigated the involvement of megestrol acetate (MA), a progesterone analog, in the stimulation of ASCs, and identifies the target receptors underlying stimulation. Mitogenic and adipogenic effects of MA were investigated in vitro, and pharmacological inhibition and small interfering (si) RNA techniques were used to identify the molecular mechanisms involved in the MA-induced stimulation of ASCs. MA significantly increased the proliferation, migration, and adipogenic differentiation of ASCs in a dose-dependent manner. Glucocorticoid receptor (GR) is highly expressed compared with other nuclear receptors in ASCs, and this receptor is phosphorylated after MA treatment. MA also upregulated genes downstream of GR in ASCs, including ANGPTL4, DUSP1, ERRF11, FKBP5, GLUL, and TSC22D3. RU486, a pharmacological inhibitor of GR, and transfection of siGR significantly attenuated MA-induced proliferation, migration, and adipogenic differentiation of ASCs. Although the adipogenic differentiation potential of MA was inferior to that of dexamethasone, MA had mitogenic effects in ASCs. Collectively, these results indicate that MA increases the proliferation, migration, and adipogenic differentiation of ASCs via GR phosphorylation. STEM CELLS TRANSLATIONAL MEDICINE 2015;4:789-799 SIGNIFICANCEMagestrol acetate (MA) increases the proliferation, migration, and adipogenic differentiation of adipose-derived stem cells (ASCs) via glucocorticoid receptor phosphorylation. Therefore, MA can be applied to increase the production yield during expansion and can be used to facilitate adipogenic differentiation of ASCs.

Wrinkle formation and abnormal pigmentation are major clinical alterations associated with skin aging. As the aim of our study was to investigate the effects of palmitoyl-KVK-L-ascorbic acid on skin aging, the anti-wrinkle and depigmentation effects of palmitoyl-KVK-L-ascorbic acid were evaluated by measuring collagen expression in dermal fibroblast cells and inhibition of melanogenesis in B16F1 cells, respectively. The anti-aging effect of palmitoyl-KVK-L-ascorbic acid cream was also evaluated against a placebo cream in a clinical trial. Our results confirmed that the expression of type Ι collagen in dermal fibroblast cells treated with palmitoyl-KVK-L-ascorbic acid (0.1-4 μg/mL) increased in a dose-dependent manner. In B16F1 cells, treatment with 20 μg/mL palmitoyl-KVK-L-ascorbic acid reduced the melanin content by approximately 20% compared to alpha-melanocyte stimulating hormone treatment. In the clinical trial, application of palmitoyl-KVK-L-ascorbic acid cream led to an improvement in skin roughness and lightness in 12 and 8 weeks, respectively. Our data show that palmitoyl-KVK-L-ascorbic acid is an effective anti-aging agent that reduces wrinkles and abnormal skin pigmentation.

Vitiligo is an acquired skin disorder characterized by the loss of pigment cells from the epidermis. We found there were abundant SENP proteins couple with the abnormal chemokines in vitiligo lesions. SUMOylation is a reversible process that is catalyzed by the enzymes and reversed by a family of Sentrin/SUMO-specific proteases(SENPs). Interferon-g (IFN-g) triggers keratinocytes for inflammation response by activating the intracellular JAKÀSTAT1 signaling. By analyzing the expression profiles of genes from SENP1-/-cells, we found the down-regulation of IFN target genes in SENP-/-cell in comparison with SENP+/+. Therefore, we proprse that SENP1 plays an important role in IFN induce signal and chemokines activation. We found the down-regulation of IFN-induced genes in SENP1-/-or SENP1-silenced KCs, and the decrease in activation of Jak2 and Stat1. SENP1-deficient KCs show defects in IFN-g signaling and type 1 T helper cells Chemokines activation. Meanwhile, we also find that SENP1 expression is induced by IFN-g. PTP1B in SENP1-deficient KCs is highly SUMOylated, which reduces PTP1B-inducedde-phosphorylation of STAT3. Activated STAT3 then suppresses STAT1 activation via SOCS3 induction in SENP1-deficient keratinocytes. Accordingly, SENP1-deficient keratinocytes show reduced ability to resist UVB. These results reveal a crucial role of SENP1-controlled STAT1 and STAT3SUMOylation balance in keratinocyte function.

Morphea is a disease of dermal inflammation and resultant sclerosis with considerable functional and cosmetic consequences. Cellular and molecular events in pathogenesis of morphea remain poorly elucidated. Preliminary studies using genome-wide gene expression analysis support the role of gamma interferon mediated pathways in the early pathogenesis of morphea. Specifically, CXCL-9 and 10, known ligands of CXCR3 receptor, were shown to be strongly upregulated in morphea. The purpose of this study is to characterize the inflammatory infiltrate in morphea and investigate cell-type specific expression of CXCR3 ligands in morphea using immunohistochemistry (IHC). Our data show that the cellular infiltrate of early inflammatory morphea lesions consists of predominantly T lymphocytes, with significantly increased CD4 positive cells and CD4:CD8 ratio of 1.8. The immune cell infiltrate of early morphea was enriched in CXCR3+ immune cells. CXCR3 co-localizes with CD4+ cells, but not CD8+ lymphocytes. We demonstrate that CXCL9 is expressed by a small subset of cells of varying morphologies within perivascular, periadnexal, and interstitial infiltrates. CXCL9 expression co-localized with CD68, but not CD20, CD34, CD123, or SMA, suggesting that dermal macrophages may secrete CXCL9 ligand. Moreover, CXCL9 expression was detected in close proximity to CXCR3 expressing immune cells. Based on our findings, we suggest that macrophage expression of CXCR3 ligands, specifically CXCL9, in morphea may enhance recruitment of CXCR-3 expressing activated T lymphocytes into the lesional skin and propagate early stages of inflammation.