Home » Efficient Differentiation of Human Pluripotent Stem Cells into Hepatocytes
Efficient Differentiation of Human Pluripotent Stem Cells into Hepatocytes
D.J.Owens, A. DeLaForest, M. Andersen, S.J. Thissen, J.R. Kettlewell and J. Aho
Secondary liver toxicity is a leading cause of commercial pharmaceutical drug recalls and compound failures during drug development. Traditional models for studying liver toxicity use hepatocytes isolated from donor livers. These primary hepatocytes can be difficult to culture, have limited proliferative capacity, and often lose functionality quickly in vitro. Hepatocyte-like cells derived from a renewable source, such as human pluripotent stem cells, are therefore a valuable resource for drug discovery research. In this study, we introduce our StemXVivo® Hepatocyte Differentiation Kit (Catalog # SC033), which efficiently directs human pluripotent stem cells through the cell fate decisions that are required for the development of functional hepatocyte-like cells. The differentiated hepatocyte-like cells were initially characterized using quantitative PCR for hepatic gene expression. We found a significant and sequential upregulation in mRNA expression for genes that encode both early and late hepatic proteins, such as Hepatocyte Nuclear Factor 4 alpha (HNF-4a), a-Fetoprotein (AFP), Transthyretin (TTR), Serpin A1 (A1AT), and Albumin. Expression of differentiation stage-specific and hepatocytespecific markers was confirmed at the protein level by immunocytochemistry and flow cytometry. Finally, pluripotent stem cell-derived hepatocyte-like cells were functionally characterized by staining for lipid and glycogen storage, quantitative analysis of albumin and urea secretion, and assessment of p450 activity. These results demonstrate that the StemXVivo® Hepatocyte Differentiation Kit easily and reproducibly generates hepatocyte-like cells from multiple pluripotent stem cell lines and can be used to provide a renewable cell source for high throughput drug discovery and screening.