Abstract: Hepatic stellate cells (HSCs), as liver-specific mesenchymal cells, play pivotal roles in liver development, regeneration, and diverse pathological processes. However, the scarcity of primary HSCs (pHSCs) and suboptimal functionality of induced HSCs (iHSCs) generated by existing methods have limited their application in biomedical modeling. In this study, we developed a de nove in vitro differentiation strategy to successfully generate functionally enhanced iHSCs capable of mimicking the liver microenvironment. Experimental validation demonstrated that these iHSCs not only exhibited core functional characteristics—including α-smooth muscle actin (α-SMA) expression, collagen secretion, and vitamin A storage—but also displayed transcriptomic profiles highly consistent with pHSCs through RNA sequencing. Notably, novel HSC-specific marker genes such as FBLN5, NID2, and SVEP1 were identified. This groundbreaking differentiation strategy achieves, for the first time, iHSCs that closely approximate pHSCs in both phenotypic and functional aspects, offering new possibilities for accurately modeling the multicellular interactive microenvironment in the liver. The generation of highly functional iHSCs not only provides an innovative platform for unraveling liver physiological regeneration mechanisms and pathological progression but also establishes a critical technical foundation for developing targeted therapeutic strategies for liver diseases.
Funding Source: National Natural Science Foundation of China (82270697), Haihe Laboratory of Cell Ecosystem (HH24KYZX0008), China Foundation For Youth Entrepreneurship and Employment -Incaier Public Welfare Fund (HH25KYHX0003)
