Improves Hepatic Differentiation from Induced Pluripotent Stem Cells by the Suppression of Epithelial-mesenchymal Transition [RNA-seq]

Background: Induced pluripotent stem cells (iPSCs) induced hepatocytes (iHeps) are widely used in modeling human liver diseases and as a potential source for cell replacement therapy. However, most iHeps are relatively immature and hard to maintain for long-term in-vitro. Results: The hepatic differentiation improved when EMT was inhibited at late stage of iHeps differentiation, and iHeps EMTi demonstrated the ability to be maintained in- vitro for an extended period up to Day 60. In-vitro analysis showed that iHeps EMTi exhibited significantly higher expression levels of hepatic functional markers, and enhanced hepatocyte functions, including lipid accumulation, glycogen storage, albumin secretion and urea acid metabolism. Moreover, the molecular profiles of iHeps EMTi are closer to those of primary human hepatocytes (PHHs). In addition, the in-vivo engraftment efficiency of iHeps EMTi was also improved as compared to iHeps alone. Conclusion: We established a robust protocol via EMT suppression to generate iHeps from human iPSCs with improved function, long-term in-vitro maintenance capacity, and enhanced repopulation efficiency. We optimized the differentiation protocol by addition of a combination of small molecules to inhibit iHeps from epithelial-mesenchymal transition (EMT, iHeps EMTi), and further characterized their function both by in-vitro and in-vivo analysis.