<p>While a liver transplant is the best therapeutic intervention for patients with severe liver failure, this life-saving procedure has significant limitations. The primary challenges are the severe shortage of organ donors, the inherent risks of major surgery, the substantial financial cost, and the need for lifelong immunosuppressive medication to prevent the body from rejecting the new organ. Accordingly, liver cell transplantation may be a promising alternative option. While aesculetin is recognized for its diverse biological properties, including anticancer, antioxidant, and anti-inflammatory effects, its potential to promote the differentiation of hepatocyte-like cells has not yet been fully investigated. Aesculetin (6,7-dihydroxycoumarin) is a bioactive compound that can facilitate the differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) into hepatocyte-like cells. In this study, we explored the effectiveness of aesculetin in directing hBM-MSCs toward a hepatic lineage. After aesculetin treatment, the hBM-MSCs exhibited increased expression of liver-specific markers such as albumin, CK-18, CK-19, CYP1A1, CYP1A2, CYP3A4, SOX17, and FOXA2, indicating successful lineage commitment. The aesculetin-treated cells demonstrated enhanced glycogen storage and increased indocyanine green uptake, demonstrating greater hepatic functionality than untreated controls. Importantly, further analysis revealed that the differentiation process promoted by aesculetin was associated with the activation of the STAT3 and STAT5 signaling pathways. Collectively, these findings underscore the pivotal role of aesculetin in promoting the hepatic differentiation of hBM-MSCs and demonstrate its potential as a key component for regenerative medicine applications in liver tissue engineering or stem cell-based therapies.</p>