Tao Liu and Shichang Zhang contributed equally to this work.
Induction of hepatocyte-like cells from mouse embryonic stem cells by lentivirus-mediated constitutive expression of Foxa2/Hnf4a
Article first published online: 12 SEP 2013
© 2013 Wiley Periodicals, Inc.
Journal of Cellular Biochemistry
Volume 114, Issue 11, pages 2531–2541, November 2013
How to Cite
Liu, T., Zhang, S., Xiang, D. and Wang, Y. (2013), Induction of hepatocyte-like cells from mouse embryonic stem cells by lentivirus-mediated constitutive expression of Foxa2/Hnf4a. J. Cell. Biochem., 114: 2531–2541. doi: 10.1002/jcb.24604
Conflict of interest: None.
- Issue published online: 12 SEP 2013
- Article first published online: 12 SEP 2013
- Accepted manuscript online: 6 JUN 2013 09:40AM EST
- Manuscript Accepted: 14 MAY 2013
- Manuscript Received: 24 JAN 2013
- National Natural Science Foundation of China. Grant Numbers: 81000182, 31070880
- Major Clinical Research Fund of Third Military Medical University
- TISSUE-ENGINEERED LIVER;
- MICRAGRAVITY BIOREACTOR;
Hepatocytes can be generated from embryonic stem cells (ESCs) using inducers such as chemical compounds and cytokines, but issues related to low differentiation efficiencies remain to be resolved. Recent work has shown that overexpression of lineage-specific transcription factors can directly cause cells phenotypic changes, including differentiation, trans-differentiation, and de-differentiation. We hypothesized that lentivirus-mediated constitutive expression of forkhead box A2 (Foxa2) and hepatocyte nuclear factor 4 alpha (Hnf4a) could promote inducing mouse ESCs to hepatocyte-likes cells. First, ESC lines that stably expressed Foxa2, Hnf4a, or Foxa2/Hnf4a were constructed via lentiviral expression vectors. Second, observations of cell morphology changes were made during the cell culture process, followed by experiments examining teratoma formation. Then, the effects of constitutive expression of Foxa2 and Hnf4a on hepatic differentiation and maturation were determined by measuring the marker gene expression levels of Albumin, α-fetoprotein, Cytokeratin18, and α1-antitrypsin. The results indicate that constitutive expression of Foxa2 and Hnf4a does not affect ESCs culture, teratoma formation, or the expression levels of the specific hepatocyte genes under autonomous differentiation. However, with some assistance from inducing factors, Foxa2 significantly increased the hepatic differentiation of ESCs, whereas the expression of Hnf4a alone or Foxa2/Hnf4a could not. Differentiated CCE-Foxa2 cells were more superior in expressing several liver-specific markers and protein, storing glycogen than differentiated CCE cells. Therefore, our method employing the transduction of Foxa2 would be a valuable tool for the efficient generation of functional hepatocytes derived from ESCs. J. Cell. Biochem. 114: 2531–2541, 2013. © 2013 Wiley Periodicals, Inc.