Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors

Abstract

The location and timing of cellular differentiation must be stringently controlled for proper organ formation. Normally, hepatocytes differentiate from hepatic progenitor cells to form the liver during development1,2. However, previous studies have shown that the hepatic program can also be activated in non-hepatic lineage cells after exposure to particular stimuli or fusion with hepatocytes3,4,5,6,7,8,9. These unexpected findings suggest that factors critical to hepatocyte differentiation exist and become activated to induce hepatocyte-specific properties in different cell types. Here, by screening the effects of twelve candidate factors, we identify three specific combinations of two transcription factors, comprising Hnf4α plus Foxa1, Foxa2 or Foxa3, that can convert mouse embryonic and adult fibroblasts into cells that closely resemble hepatocytes in vitro. The induced hepatocyte-like (iHep) cells have multiple hepatocyte-specific features and reconstitute damaged hepatic tissues after transplantation. The generation of iHep cells may provide insights into the molecular nature of hepatocyte differentiation and potential therapies for liver diseases.

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Figure 1: Generation of iHep cells from MEF cultures and hepatocyte-specific properties in iHep cells.
Figure 2: Hepatic functions in iHep cells.
Figure 3: iHep cells reconstitute hepatic tissues and support hepatic function in vivo.

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Gene Expression Omnibus

Data deposits

Our microarray data have been uploaded to the Gene Expression Omnibus database, accession number GSE29725.

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Acknowledgements

We thank A. Iwama, H. Miyoshi and R. M. Tanguay for sharing reagents, F. Costantini for providing the R26RYFP mice and E. Gunshima, A. Kaneyuki and H. Kuboyama for excellent technical assistance. This work was supported in part by the Program for Improvement of the Research Environment for Young Researchers from the Special Coordination Funds for Promoting Science and Technology commissioned by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, a Grant-in-Aid for Scientific Research from the MEXT of Japan and the Precursory Research for Embryonic Science and Technology Program of the Japan Science and Technology Agency.

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A.S. designed the study and wrote the paper. S.S. and A.S. performed experiments and analysed and interpreted the data.

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Correspondence to Atsushi Suzuki.

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The authors declare no competing financial interests.

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This file contains a Supplementary Discussion, Supplementary References, Supplementary Figures 1-23 with legends and Supplementary Table 1. (PDF 30029 kb)

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Sekiya, S., Suzuki, A. Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors. Nature 475, 390–393 (2011). https://doi.org/10.1038/nature10263

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