In vitro modeling of liver fibrosis with 3D co‐culture system using a novel human hepatic stellate cell line

Abstract

AbstractHepatic stellate cells (HSCs) play an important role in liver fibrosis; however, owing to the heterogeneity and limited supply of primary HSCs, the development of in vitro liver fibrosis models has been impeded. In this study, we established and characterized a novel human HSC line (LSC‐1), and applied it to various types of three‐dimensional (3D) co‐culture systems with differentiated HepaRG cells. Furthermore, we compared LSC‐1 with a commercially available HSC line on conventional monolayer culture. LSC‐1 exhibited an overall upregulation of the expression of fibrogenic genes along with increased levels of matrix and adhesion proteins, suggesting a myofibroblast‐like or transdifferentiated state. However, activated states reverted to a quiescent‐like phenotype when cultured in different 3D culture formats with a relatively soft microenvironment. Additionally, LSC‐1 exerted an overall positive effect on co‐cultured differentiated HepaRG, which significantly increased hepatic functionality upon long‐term cultivation compared with that achieved with other HSC line. In 3D spheroid culture, LSC‐1 exhibited enhanced responsiveness to transforming growth factor beta 1 exposure that is caused by a different matrix‐related protein expression mechanism. Therefore, the LSC‐1 line developed in this study provides a reliable candidate model that can be used to address unmet needs, such as development of antifibrotic therapies.