Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo

David C. Hay, Debiao Zhao, Judy Fletcher, Zoe A. Hewitt, Doris McLean, Alai Urruticoechea-Uriguen, James R. Black, Cliff Elcombe, James A. Ross, Roland Wolf, Wei Cui

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    297 Citations (Scopus)

    Abstract

    The potential to differentiate human embryonic stem cells (hESCs) in vitro to provide an unlimited source of human hepatocytes for use in biomedical research, drug discovery, and the treatment of liver diseases holds great promise. Here we describe a three-stage process for the efficient and reproducible differentiation of hESCs to hepatocytes by priming hESCs towards definitive endoderm with activin A and sodium butyrate prior to further differentiation to hepatocytes with dimethyl sulfoxide, followed by maturation with hepatocyte growth factor and oncostatin M. We have demonstrated that differentiation of hESCs in this process recapitulates liver development in vivo: following initial differentiation, hESCs transiently express characteristic markers of the primitive streak mesendoderm before turning to the markers of the definitive endoderm; with further differentiation, expression of hepatocyte progenitor cell markers and mature hepatocyte markers emerged sequentially. Furthermore, we have provided evidence that the hESC-derived hepatocytes are able to carry out a range of hepatocyte functions: storage of glycogen, and generation and secretion of plasma proteins. More importantly, the hESC-derived hepatocytes express several members of cytochrome P450 isozymes, and these P450 isozymes are capable of converting the substrates to metabolites and respond to the chemical stimulation. Our results have provided evidence that hESCs can be differentiated efficiently in vitro to functional hepatocytes, which may be useful as an in vitro system for toxicity screening in drug discovery.

    Original languageEnglish
    Pages (from-to)894-902
    Number of pages9
    JournalStem Cells
    Volume26
    Issue number4
    DOIs
    Publication statusPublished - Apr 2008

    Keywords

    • hepatocyte differentiation
    • human embryonic stem cell
    • cytochrome P450
    • liver development
    • FUNCTIONAL HEPATIC CELLS
    • DEFINITIVE ENDODERM
    • TRANSCRIPTIONAL CONTROL
    • HUMAN BLASTOCYSTS
    • SODIUM-BUTYRATE
    • ACTIVIN-A
    • CULTURE
    • EXPRESSION
    • LINES
    • VITRO

    Cite this

    Hay, D. C., Zhao, D., Fletcher, J., Hewitt, Z. A., McLean, D., Urruticoechea-Uriguen, A., Black, J. R., Elcombe, C., Ross, J. A., Wolf, R., & Cui, W. (2008). Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo. Stem Cells, 26(4), 894-902. https://doi.org/10.1634/stemcells.2007-0718