Induction of functional hepatocyte-like cells from mouse fibroblasts by defined factors

Abstract

The generation of functional hepatocytes independent of donor liver organs is of great therapeutic interest with regard to regenerative medicine and possible cures for liver disease1. Induced hepatic differentiation has been achieved previously using embryonic stem cells or induced pluripotent stem cells2,3,4,5,6,7,8. Particularly, hepatocytes generated from a patient’s own induced pluripotent stem cells could theoretically avoid immunological rejection. However, the induction of hepatocytes from induced pluripotent stem cells is a complicated process that would probably be replaced with the arrival of improved technology. Overexpression of lineage-specific transcription factors directly converts terminally differentiated cells into some other lineages9,10,11,12, including neurons13, cardiomyocytes14 and blood progenitors15; however, it remains unclear whether these lineage-converted cells could repair damaged tissues in vivo. Here we demonstrate the direct induction of functional hepatocyte-like (iHep) cells from mouse tail-tip fibroblasts by transduction of Gata4, Hnf1α and Foxa3, and inactivation of p19Arf. iHep cells show typical epithelial morphology, express hepatic genes and acquire hepatocyte functions. Notably, transplanted iHep cells repopulate the livers of fumarylacetoacetate-hydrolase-deficient (Fah−/−) mice and rescue almost half of recipients from death by restoring liver functions. Our study provides a novel strategy to generate functional hepatocyte-like cells for the purpose of liver engineering and regenerative medicine.

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Figure 1: Three transcription factors induce hepatic conversion of tail-tip fibroblasts.
Figure 2: Characterization of iHep cells in vitro.
Figure 3: iHep cell transplantation rescues Fah-deficient mice.
Figure 4: iHep cells restore liver functions of Fah −/− Rag2 −/− mice.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data have been deposited in the Gene Expression Omnibus database under accession number GSE23635.

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Acknowledgements

We would like to thank X. Chen, J. Cen, L. Zhang and J. Yuan for technical support, and F. Tang and D. Li for comments. The laboratory of L.H. is funded by the National Science Foundation of China (91019014) and the Chinese Academy of Sciences (KSCX2-YW-R-241, XDA01010402 and the Hundred Talents Program). The laboratory of X.W. is supported by the National Science Foundation of China (30801115), Chinese National 863 Plan Project (2006AA02Z474) and National Key Basic Research and Development Program of China (2007CB947102 and 2009CB941100).

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L.H. conceived the project. P.H. performed most of the experiments. S.J. analysed the in vitro functions of iHep cells. H.S. analysed gene expression of iHep cells. L.H., X.W., P.H. and Z.H. designed the experiments for characterizing in vivo functions of iHep cells. P.H., Z.H., D.X., C.L. and Y.H. performed the in vivo experiments. L.H. and P.H. analysed the data. L.H., P.H. and X.W. wrote the manuscript.

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Correspondence to Lijian Hui.

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The authors declare no competing financial interests.

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This file contains Supplementary Tables 1-3 and Supplementary Figures 1-13 with legends. (PDF 2651 kb)

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Huang, P., He, Z., Ji, S. et al. Induction of functional hepatocyte-like cells from mouse fibroblasts by defined factors. Nature 475, 386–389 (2011). https://doi.org/10.1038/nature10116

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