Ye Di, Ding Min, Song Yu-Mu, Meng Heng-Xing, Chen Wen-Hao, Ge Jian-Yun, Zheng Yun-Wen
Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, and South China, Institute of Large Animal Models for Biomedicine, School of Pharmacy and FoodEngineering, Wuyi University, Jiangmen, 529020, Guangdong, China.
Haihe Laboratory of Cell Ecosystem, Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin, 300020, China.
Sci Rep. 2025 Jul 1;15(1):22338. doi: 10.1038/s41598-025-09026-7.
Hepatic stellate cells (HSCs) are liver-specific mesenchymal cells that play a crucial role in liver formation and regeneration, as well as in different pathological diseases. However, the limited source of primary HSCs (pHSCs) and the suboptimal functionality of induced HSCs (iHSCs) by existing methods restrict their application in biomedical modeling. We developed a de novo differentiation method to generate iHSCs under simulated liver microenvironment in vitro, thereby enhancing the function of the differentiated cells. These iHSCs exhibited key HSC functions, including the expression of α-smooth muscle actin, collagen, and the capability to store Vitamin A. RNA sequencing further revealed that the present iHSCs converged more closely to pHSCs with very similar transcriptional profile compared to the established conventional induction. Additionally, the novel HSC-specific marker genes, FBLN5, NID2, and SVEP1 were identified by RNA sequencing and gene expression assay. In conclusion, our novel differentiation approach enables the generation of iHSCs with phenotypic and functional traits similar to those of pHSCs. The generation of highly functional iHSCs may make it more feasible to accurately simulate the liver-specific multicellular microenvironments, thus providing new perspectives on the modeling of physiological regenerative processes and disease progression in the liver, as well as useful tools for creating of new therapeutic strategies.
肝星状细胞（HSCs）是肝脏特异性间充质细胞，在肝脏形成、再生以及不同病理疾病中发挥关键作用。然而，原代肝星状细胞（pHSCs）来源有限，且现有方法诱导的肝星状细胞（iHSCs）功能欠佳，限制了它们在生物医学建模中的应用。我们开发了一种从头分化方法，在体外模拟肝脏微环境下生成iHSCs，从而增强分化细胞的功能。这些iHSCs表现出关键的肝星状细胞功能，包括α平滑肌肌动蛋白、胶原蛋白的表达以及储存维生素A的能力。RNA测序进一步显示，与已确立的传统诱导方法相比，目前的iHSCs与pHSCs的转录谱更为相似，更接近pHSCs。此外，通过RNA测序和基因表达分析鉴定了新的肝星状细胞特异性标记基因FBLN5、NID2和SVEP1。总之，我们的新型分化方法能够生成具有与pHSCs相似表型和功能特征的iHSCs。高功能iHSCs的生成可能使准确模拟肝脏特异性多细胞微环境变得更可行，从而为肝脏生理再生过程和疾病进展的建模提供新视角，以及为创建新的治疗策略提供有用工具。
