Abstract
The human liver is a vital organ within the body and plays a major role in normal homeostasis. The “work horse” of the liver, termed the “hepatocyte,” is estimated to make up approximately 70–80% of the liver’s mass. Therefore, the study of hepatocyte biology has an important role to play in medicine and the drug discovery process. At present the routine use of human primary hepatocytes is limited due to poor supply and their loss of function upon isolation. Therefore, additional and renewable sources of hepatocytes are being sought. Rodent hepatocytes have been utilised for many years, and although informative, they possess significant limitations and do not accurately extrapolate to human liver. To overcome the issue of cell viability, several groups have tried to generate immortalised hepatocytes; however, the derivative cells exhibit dramatic decreases in function and karyotypic instability over prolonged culture. It has therefore been necessary to find an alternative source of hepatocytes and efficient methods for deriving hepatic endoderm from stem cells in vitro. We have employed human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) to derive human hepatic endoderm (HE). hESCs and iPSCs represent scalable and highly efficient resources with which to generate human HE in vitro, and hESC-derived HE will be the focus of this chapter.
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References
Wege H, Chui MS, Le HT, et al. (2003). In vitro expansion of human hepatocytes is restricted by telomere-dependent replicative aging. Cell Transplant 12: 897–906.
Cai J, Ito M, Westerman KA, et al. (2000). Construction of a non-tumorigenic rat hepatocyte cell line for transplantation: reversal of hepatocyte immortalization by site-specific excision of the SV40 T antigen. J Hepatol 33: 701–708.
Allain JE, Dagher I, Mahieu-Caputo D, et al. (2002). Immortalization of a primate bipotent epithelial liver stem cell. Proc Natl Acad Sci USA 99: 3639–3644.
Delgado JP, Parouchev A, Allain JE, et al. (2005). Long-term controlled immortalization of a primate hepatic progenitor cell line after Simian virus 40 T-Antigen gene transfer. Oncogene 24: 541–551.
Turpeinen M, Tolonen A, Chesne C, et al. (2009). Functional expression, inhibition and induction of CYP enzymes in HepaRG cells. Toxicol In Vitro 23: 748–753.
Yoshitomi S, Ikemoto K, Takahashi J, et al. (2001). Establishment of the transformants expressing human cytochrome P450 subtypes in HepG2, and their applications on drug metabolism and toxicology. Toxicol In Vitro 15: 245–256.
Herrera MB, Bruno S, Buttiglieri S, et al. (2006). Isolation and characterization of a stem cell population from adult human liver. Stem Cells 24: 2840–2850.
Lazaro CA, Rhim JA, Yamada Y, and Fausto N. (1998). Generation of hepatocytes from oval cell precursors in culture. Cancer Res 58: 5514–5522.
Rogler LE. (1997) Selective bipotential differentiation of mouse embryonic hepatoblasts in vitro. Am J Pathol 150: 591–602.
Reubinoff BE, Pera MF, Fong CY, et al. (2000). Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18: 399–404.
Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. (1998). Embryonic stem cell lines derived from human blastocysts. Science 282: 1145–1147.
Takahashi K, and Yamanaka S. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126: 663–676.
Dalgetty DM, Medine C, Iredale JP, et al. (2009). Progress and Future Challenges in Stem Cell-Derived Liver Technologies. American Journal of Physiology – Gastrointestinal and Liver Physiology. 2009. 297(2):G241–248.
Greenhough S, Medine CN, and Hay DC. (2010). Pluripotent Stem Cell Derived Hepatocyte Like Cells and their Potential in ToxicityScreening. Toxicology 278: 250–255
Hay DC. (2010) Cadaveric Hepatocytes Repopulate Diseased Livers: Life After Death. Gastroenterology 139: 715–716
Medine CN, Greenhough S, and Hay DC. (2010). Role of stem-cell-derived hepatic endoderm in human drug discovery. Biochem. Soc. Trans 38: 1033–1036
Agarwal S, Holton KL, Lanza R, et al. (2008). Efficient differentiation of functional hepatocytes from human embryonic stem cells. Stem Cells 26: 1117–1127.
Basma H, Soto-Gutierrez A, Yannam G, et al. (2009). Differentiation and Transplantation of Human Embryonic Stem Cell-Derived Hepatocytes. Gastroenterology 136: 990–999.
Cai J, Zhao Y, Liu Y, et al. (2007). Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology 45: 1229–1239.
Duan Y, Catana A, Meng Y, et al. (2007). Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo. Stem Cells 25: 3058–3068.
Fletcher J, Cui W, Samuel K, et al. (2008). The inhibitory role of stromal cell mesenchyme on human embryonic stem cell hepatocyte differentiation is overcome by Wnt3a treatment. Cloning Stem Cells 10: 331–339.
Hay DC, Zhao D, Ross A, et al. (2007). Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. Cloning Stem Cells 9: 51–62.
Hay DC, Zhao D, Fletcher J, et al. (2008). Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo. Stem Cells 26: 894–902.
Hay DC, Fletcher J, Payne C, et al. (2008). Highly efficient differentiation of hESCs to functional hepatic endoderm requires ActivinA and Wnt3a signaling. Proc Natl Acad Sci USA 105: 12301–12306.
Sullivan GJ, Hay DC, Park IH, et al. (2010). Generation of functional human hepatic endoderm from human induced pluripotent stem cells. Hepatology 51: 329–335.
Hannoun Z, Fletcher J, Greenhough S, et al.(2010). The Comparison between Conditioned Media and Serum-Free Media in Human Embryonic Stem Cell Culture and Differentiation, Cellular Reprogramming, 12:2 133–140.
Hay DC, Pernagallo S, Diaz-Mochon JJ, et al (2011). Unbiased Screening of Polymer Libraries to Define Novel Substrates for Functional Hepatocytes with Inducible Drug Metabolism. Stem Cell Research, 6: 92–101.
Payne C.M, Samuel K, Pryde A, et al (2011).Persistence of Functional Hepatocyte Like Cells in Immune Compromised Mice. Liver International, 31(2):254–62.
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© 2011 Humana Press
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Medine, C.N., Hannoun, Z., Greenhough, S., Payne, C.M., Fletcher, J., Hay, D.C. (2011). Deriving Metabolically Active Hepatic Endoderm from Pluripotent Stem Cells. In: Ye, K., Jin, S. (eds) Human Embryonic and Induced Pluripotent Stem Cells. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-61779-267-0_27
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DOI: https://doi.org/10.1007/978-1-61779-267-0_27
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