Stem Cell Reviews and Reports

, Volume 13, Issue 4, pp 542–551 | Cite as

Hepatocyte Nuclear Factor 4 Alpha Promotes Definitive Endoderm Differentiation from Human Induced Pluripotent Stem Cells

  • Morifumi Hanawa
  • Kazuo Takayama
  • Fuminori Sakurai
  • Masashi Tachibana
  • Hiroyuki Mizuguchi


Hepatocyte nuclear factor 4 alpha (HNF4α) is a key transcription factor for liver development. Although HNF4α is necessary for hepatoblast differentiation, the function of HNF4α before the hepatoblast differentiation, such as in definitive endoderm differentiation, is not well known. In addition, it is known that there are nine HNF4α isoforms, but the expression and function of each HNF4α isoform during the definitive endoderm differentiation is also not clear. In this study, we examined the expression pattern of HNF4α and its functions in the definitive endoderm differentiation from human induced pluripotent stem (iPS) cells. We found that the HNF4α-1D isoform expression levels were significantly increased during the definitive endoderm differentiation, while the HNF4α-1A isoform expression levels did not change. Therefore, we further examined the function of the HNF4α-1D isoform in definitive endoderm differentiation. HNF4α-1D overexpression or knockdown was found to promote or prevent the definitive endoderm differentiation, respectively. Interestingly, Lefty1 was directly regulated by HNF4α-1D, and Lefty1 knockdown also prevented the definitive endoderm differentiation. These results suggest that HNF4α-1D promotes definitive endoderm differentiation through the regulation of Lefty1. To our knowledge, this is the first report to clarify the expression pattern and function of HNF4α during the definitive endoderm differentiation.


HNF4α Definitive endoderm Human iPS cells Lefty 1 Differentiation 



We thank Ms. Yasuko Hagihara, Ms. Natsumi Mimura, and Ms. Ayaka Sakamoto for their excellent technical support. This research was supported by the Keihanshin Consortium for Fostering the Next Generation of Global Leaders in Research (K-CONNEX), established by Human Resource Development Program for Science and Technology, MEXT. This research was also supported by the grants (15bk0104005h0003) from Japan Agency for Medical Research and development, AMED.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no competing financial interests.

Supplementary material

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Table S1 (DOCX 14 kb)
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Table S2 (DOCX 13 kb)
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Table S3 (DOCX 13 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Morifumi Hanawa
    • 1
  • Kazuo Takayama
    • 1
    • 2
    • 3
    • 4
  • Fuminori Sakurai
    • 1
  • Masashi Tachibana
    • 1
  • Hiroyuki Mizuguchi
    • 1
    • 4
    • 5
    • 6
  1. 1.Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical SciencesOsaka UniversityOsakaJapan
  2. 2.The Keihanshin Consortium for Fostering the Next Generation of Global Leaders in Research (K-CONNEX)Kyoto UniversityKyotoJapan
  3. 3.PRESTO, Japan Science and Technology AgencySaitamaJapan
  4. 4.Laboratory of Hepatocyte Differentiation, National Institute of Biomedical Innovation, Health and NutritionOsakaJapan
  5. 5.iPS Cell-Based Research Project on Hepatic Toxicity and Metabolism, Graduate School of Pharmaceutical SciencesOsaka UniversityOsakaJapan
  6. 6.Global Center for Medical Engineering and InformaticsOsaka UniversityOsakaJapan

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