Enhancement of hepatocyte differentiation from human embryonic stem cells by Chinese medicine Fuzhenghuayu

Jiamei Chen,Wei Gao,Mark A Zern,Chenghai Liu,Benjamin Tschudy-Seney,Yuyou Duan,Xiaocui Ma,Ping Liu,Ping Zhou

doi:10.1038/srep18841

Abstract

Chinese medicine, Fuzhenghuayu (FZHY), appears to prevent fibrosis progression and improve liver function in humans. Here we found that FZHY enhanced hepatocyte differentiation from human embryonic stem cells (hESC). After treatment with FZHY, albumin expression was consistently increased during differentiation and maturation process, and expression of metabolizing enzymes and transporter were also increased. Importantly, expression of mesenchymal cell and cholangiocyte marker was significantly reduced by treatment with FZHY, indicating that one possible mechanism of FZHY’s role is to inhibit the formation of mesenchymal cells and cholangiocytes. Edu-labelled flow cytometric analysis showed that the percentage of the Edu positive cells was increased in the treated cells. These results indicate that the enhanced proliferation involved hepatocytes rather than another cell type. Our investigations further revealed that these enhancements by FZHY are mediated through activation of canonical Wnt and ERK pathways and inhibition of Notch pathway. Thus, FZHY not only promoted hepatocyte differentiation and maturation, but also enhanced hepatocyte proliferation. These results demonstrate that FZHY appears to represent an excellent therapeutic agent for the treatment of liver fibrosis, and that FZHY treatment can enhance our efforts to generate mature hepatocytes with proliferative capacity for cell-based therapeutics and for pharmacological and toxicological studies.

Highlights

Hepatocytes exhibit in vivo function shown by engrafting and proliferation in mouse livers[16]
In our screening tests with different concentrations of FZHY and the addition of FZHY at different time points during the differentiation process, we found that human embryonic stem cells (hESC)-derived hepatocyte differentiation and maturation could be promoted at the concentration of 50 and 100 μ g/ml FZHY and the addition times at days 8 and 20 for 6 days
The differentiating cells were treated with FZHY between days 8–14, whereas FZHY was added between days 20–26 during the maturation process (Fig. 1A)

Summary

Introduction

Hepatocytes exhibit in vivo function shown by engrafting and proliferation in mouse livers[16]. QPCR results revealed that these genes were highly expressed during both differentiation and maturation processes in the treated cells when compared to cells without treatment (Fig. 2B,C).

Results

Conclusion