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Rapid generation of mature hepatocyte‐like cells from human induced pluripotent stem cells by an efficient three‐step protocol
Author(s) -
Chen YuFan,
Tseng ChienYu,
Wang HseiWei,
Kuo HungChih,
Yang Vincent W.,
Lee Oscar K.
Publication year - 2012
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.24790
Subject(s) - induced pluripotent stem cell , biology , hepatocyte , microbiology and biotechnology , transplantation , stem cell , cancer research , hepatocyte growth factor , embryonic stem cell , medicine , biochemistry , in vitro , receptor , gene
Liver transplantation is the only definitive treatment for end‐stage cirrhosis and fulminant liver failure, but the lack of available donor livers is a major obstacle to liver transplantation. Recently, induced pluripotent stem cells (iPSCs) derived from the reprogramming of somatic fibroblasts, have been shown to resemble embryonic stem (ES) cells in that they have pluripotent properties and the potential to differentiate into all cell lineages in vitro , including hepatocytes. Thus, iPSCs could serve as a favorable cell source for a wide range of applications, including drug toxicity testing, cell transplantation, and patient‐specific disease modeling. Here, we describe an efficient and rapid three‐step protocol that is able to rapidly generate hepatocyte‐like cells from human iPSCs. This occurs because the endodermal induction step allows for more efficient and definitive endoderm cell formation. We show that hepatocyte growth factor (HGF), which synergizes with activin A and Wnt3a, elevates the expression of the endodermal marker Foxa2 (forkhead box a2) by 39.3% compared to when HGF is absent (14.2%) during the endodermal induction step. In addition, iPSC‐derived hepatocytes had a similar gene expression profile to mature hepatocytes. Importantly, the hepatocyte‐like cells exhibited cytochrome P450 3A4 (CYP3A4) enzyme activity, secreted urea, uptake of low‐density lipoprotein (LDL), and possessed the ability to store glycogen. Moreover, the hepatocyte‐like cells rescued lethal fulminant hepatic failure in a nonobese diabetic severe combined immunodeficient mouse model. Conclusion : We have established a rapid and efficient differentiation protocol that is able to generate functional hepatocyte‐like cells from human iPSCs. This may offer an alternative option for treatment of liver diseases. (H epatology 2012)