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Identification of small molecules for human hepatocyte expansion and iPS differentiation

Nature Chemical Biology volume 9, pages 514–520 (2013)Cite this article

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Abstract

Cell-based therapies hold the potential to alleviate the growing burden of liver diseases. Such therapies require human hepatocytes, which, within the stromal context of the liver, are capable of many rounds of replication. However, this ability is lost ex vivo, and human hepatocyte sourcing has limited many fields of research for decades. Here we developed a high-throughput screening platform for primary human hepatocytes to identify small molecules in two different classes that can be used to generate renewable sources of functional human hepatocytes. The first class induced functional proliferation of primary human hepatocytes in vitro. The second class enhanced hepatocyte functions and promoted the differentiation of induced pluripotent stem cell–derived hepatocytes toward a more mature phenotype than what was previously obtainable. The identification of these small molecules can help address a major challenge affecting many facets of liver research and may lead to the development of new therapeutics for liver diseases.

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Figure 1: High-throughput identification of small molecules that induce proliferation and enhance the functions of primary human hepatocytes.
Figure 2: Expansion of primary human hepatocytes.
Figure 3: Functional enhancement of human primary hepatocytes and iHep cells.
Figure 4: Maturation of human iHep cells.

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Acknowledgements

This research was funded by a Broad Institute Scientific Planning and Allocation of Resources Committee grant (S.N.B.), US National Institutes of Health (NIH) grants NIH R01-DK065152 (S.N.B.), NIH R01-DK56966 (S.N.B.) and NIH R01 GM089652 (A.E.C.), National Science Foundation CAREER award DBI 1148823 (A.E.C.) and National Institute of Diabetes and Digestive and Kidney Diseases grants DK085445 (W.G.), DK55743 (S.A.D.), DK087377 (S.A.D.), HL094857 (S.A.D.) and HG006398 (S.A.D.). S.N.B. is a Howard Hughes Medical Institute investigator. We thank H. Fleming for help with manuscript preparation, T. Golub and R. Gould for discussions and H. Green (Harvard University) for providing J2-3T3 fibroblasts. The authors dedicate this paper to the memory of Officer Sean Collier, for his caring service to the MIT community and his sacrifice.

Author information

Authors and Affiliations

  1. Harvard–Massachusetts Institute of Technology (MIT) Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA

    Jing Shan, Robert E Schwartz & Sangeeta N Bhatia

  2. Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Robert E Schwartz & Sangeeta N Bhatia

  3. Chemical Biology Platform, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Nathan T Ross

  4. Imaging Platform, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    David J Logan & Anne E Carpenter

  5. Cancer Program, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    David Thomas

  6. Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

    Stephen A Duncan

  7. Harvard Medical School, Boston, Massachusetts, USA

    Trista E North & Wolfram Goessling

  8. Harvard Stem Cell Institute, Cambridge, Massachusetts, USA

    Trista E North & Wolfram Goessling

  9. Department of Pathology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA

    Trista E North

  10. Genetics Division, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Wolfram Goessling

  11. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Sangeeta N Bhatia

  12. Institute for Medical Engineering and Science, MIT, Cambridge, Massachusetts, USA

    Sangeeta N Bhatia

  13. Department of Electrical Engineering and Computer Science, MIT, Cambridge, Massachusetts, USA

    Sangeeta N Bhatia

  14. David H. Koch Institute for Integrative Cancer Research, MIT, Cambridge, Massachusetts, USA

    Sangeeta N Bhatia

  15. Howard Hughes Medical Institute, Massachusetts, USA

    Sangeeta N Bhatia

Authors
  1. Jing Shan
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Contributions

J.S. and S.N.B. designed all experiments. J.S. performed and analyzed all experiments. D.J.L. helped analyze image-based experiments. N.T.R. helped perform and analyze screening experiments and analyzed chemical characterization data. R.E.S. helped design, perform and analyze iPS experiments. D.T. developed and implemented the Luminex system. T.E.N., S.A.D., W.G. and A.E.C. contributed reagents and expertise. J.S., S.N.B. and N.T.R. wrote the manuscript. S.N.B. supervised the project.

Corresponding author

Correspondence to Sangeeta N Bhatia.

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Shan, J., Schwartz, R., Ross, N. et al. Identification of small molecules for human hepatocyte expansion and iPS differentiation. Nat Chem Biol 9, 514–520 (2013). https://doi.org/10.1038/nchembio.1270

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