Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Cell & Tissue Engineering, Cell Biology, DEFINITIVE ENDODERM, DIFFERENTIATION, SPECIFICATION, REGENERATION, HEPATOCYTES, COMMITMENT, EXPRESSION, RECEPTOR, MARKER, INJURY, differentiation, disease modeling, hepatic stellate cells, in vitro liver model, liver fibrosis, liver spheroids, non-parenchymal cells, organoids, pluripotent stem cells, toxicity assessment, Cell Differentiation, Cells, Cultured, Coculture Techniques, Female, Hepatic Stellate Cells, Humans, Infant, Newborn, Liver Cirrhosis, Male, Models, Biological, Pluripotent Stem Cells, Thioacetamide, Wound Healing, EU-ToxRisk - 681002;info:eu-repo/grantAgreement/EC/H2020/681002, 06 Biological Sciences, 11 Medical and Health Sciences, Developmental Biology, 31 Biological sciences, 32 Biomedical and clinical sciences

Abstract:

The development of complex in vitro hepatic systems and artificial liver devices has been hampered by the lack of reliable sources for relevant cell types, such as hepatic stellate cells (HSCs). Here we report efficient differentiation of human pluripotent stem cells into HSC-like cells (iPSC-HSCs). iPSC-HSCs closely resemble primary human HSCs at the transcriptional, cellular, and functional levels and possess a gene expression profile intermediate between that of quiescent and activated HSCs. Functional analyses revealed that iPSC-HSCs accumulate retinyl esters in lipid droplets and are activated in response to mediators of wound healing, similar to their in vivo counterparts. When maintained as 3D spheroids with HepaRG hepatocytes, iPSC-HSCs exhibit a quiescent phenotype but mount a fibrogenic response and secrete pro-collagen in response to known stimuli and hepatocyte toxicity. Thus, this protocol provides a robust in vitro system for studying HSC development, modeling liver fibrosis, and drug toxicity screening.