These authors contributed equally to this article.
Directed differentiation of human embryonic stem cells into functional hepatic cells†
Article first published online: 26 APR 2007
Copyright © 2007 American Association for the Study of Liver Diseases
Volume 45, Issue 5, pages 1229–1239, May 2007
How to Cite
Cai, J., Zhao, Y., Liu, Y., Ye, F., Song, Z., Qin, H., Meng, S., Chen, Y., Zhou, R., Song, X., Guo, Y., Ding, M. and Deng, H. (2007), Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology, 45: 1229–1239. doi: 10.1002/hep.21582
Potential conflict of interest: Nothing to report.
- Issue published online: 26 APR 2007
- Article first published online: 26 APR 2007
- Manuscript Accepted: 7 DEC 2006
- Manuscript Received: 1 SEP 2006
- Bill & Melinda Gates Foundation. Grant Number: 37871
- Science and Technology Plan of the Beijing Municipal Government. Grant Number: H020220050290
- Beijing Nature Science Foundation. Grant Number: SO51003
The differentiation capacity of human embryonic stem cells (hESCs) holds great promise for therapeutic applications. We report a novel three-stage method to efficiently direct the differentiation of human embryonic stem cells into hepatic cells in serum-free medium. Human ESCs were first differentiated into definitive endoderm cells by 3 days of Activin A treatment. Next, the presence of fibroblast growth factor-4 and bone morphogenetic protein-2 in the culture medium for 5 days induced efficient hepatic differentiation from definitive endoderm cells. Approximately 70% of the cells expressed the hepatic marker albumin. After 10 days of further in vitro maturation, these cells expressed the adult liver cell markers tyrosine aminotransferase, tryptophan oxygenase 2, phosphoenolpyruvate carboxykinase (PEPCK), Cyp7A1, Cyp3A4 and Cyp2B6. Furthermore, these cells exhibited functions associated with mature hepatocytes including albumin secretion, glycogen storage, indocyanine green, and low-density lipoprotein uptake, and inducible cytochrome P450 activity. When transplanted into CCl4 injured severe combined immunodeficiency mice, these cells integrated into the mouse liver and expressed human alpha-1 antitrypsin for at least 2 months. In addition, we found that the hESC-derived hepatic cells were readily infected by human immunodeficiency virus-hepatitis C virus pseudotype viruses. Conclusion: We have developed an efficient way to direct the differentiation of human embryonic stem cells into cells that exhibit characteristics of mature hepatocytes. Our studies should facilitate searching the molecular mechanisms underlying human liver development, and form the basis for hepatocyte transplantation and drug tests. (HEPATOLOGY 2007;45:1229–1239.)