Liver developmental microenvironment promotes iHSC generation from human iPSCs

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 iHSC 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.