Abstract
Human embryonic stem cells (HESCs) are pluripotent cells having a self-renewal capacity. These unique characteristics of HESCs allow them to be an unlimited source of cells that was shown to differentiate into many cell types, among them hepatocytes. The creation of hepatocytes in culture will allow us to further understand the mechanisms involved in the embryogenesis of hepatocytes in humans and to study pathologies related to aberrant differentiation of these cells. The resultant hepatocytes may serve as a source of cells for transplantation and as cells for toxicological studies and drug screening. In the past 10 years, since the derivation of HESCs, various protocols for the differentiation of HESCs to hepatic-like cells were published. In this chapter we detail our protocol for differentiating HESCs into hepatic-like cells through embryoid bodies. We further describe the method for the genetic labeling of the hepatic-like cells derived from the HESCs and their isolation by fluorescence-activated cell sorter. We also summarize the published protocols for differentiation of HESCs into hepatic-like cells.
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References
Reubinoff, B.E., Pera, M.F., Fong, C.Y., Trounson, A., and Bongso, A. (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechnol. 18, 399–404.
Thomson, J.A. et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.
Itskovitz-Eldor, J. et al. (2000) Differentiation of human embyronic stem cells into embryoid bodies comprising the three embryonic germ layers. Mol. Med. 6, 88–95.
Lavon, N. and Benvenisty, N. (2005) Study of hepatocyte differentiation using embryonic stem cells. J. Cell Biochem. 96, 1193–1202.
Schleger, C., Krebsfaenger, N., Kalkuhl, A., Bader, R., and Singer, T. (2001) Innovative cell culture methods in drug development. Altex 18, 5–8.
Schuldiner, M., Yanuka, O., Itskovitz-Eldor, J., Melton, D.A., and Benvenisty, N. (2000) Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. Proc. Natl. Acad. Sci. USA 97, 11307–11312.
Lavon, N., Yanuka, O., and Benvenisty, N. (2004) Differentiation and isolation of hepatic-like cells from human embryonic stem cells. Differentiation 72, 230–238.
Agarwal, S., Holton, K.L., and Lanza, R. (2008) Efficient differentiation of functional hepatocytes from human embryonic stem cells. Stem Cells 26, 1117–1127.
Baharvand, H., Hashemi, S.M., Kazemi Ashtiani, S., and Farrokhi, A. (2006) Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro. Int. J. Dev. Biol. 50, 645–652.
Baharvand, H., Hashemi, S.M., and Shahsavani, M. (2008) Differentiation of human embryonic stem cells into functional hepatocyte-like cells in a serum-free adherent culture condition. Differentiation 76, 465–477.
Cai, J. et al. (2007) Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology 45, 1229–1239.
Chiao, E. et al. (2008) Isolation and transcriptional profiling of purified hepatic cells derived from human embryonic stem cells. Stem Cells 26, 2032–2041.
Duan, 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.
Ek, M. et al. (2007) Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells. Biochem. Pharmacol. 74, 496–503.
Hay, D.C. 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, D.C. et al. (2007) Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. Cloning Stem Cells 9, 51–62.
Ishii, T. et al. (2008) Effects of extracellular matrixes and growth factors on the hepatic differentiation of human embryonic stem cells. Am. J. Physiol. Gastrointest. Liver Physiol. 295, G313–G321.
Rambhatla, L., Chiu, C.P., Kundu, P., Peng, Y., and Carpenter, M.K. (2003) Generation of hepatocyte-like cells from human embryonic stem cells. Cell Transplant. 12, 1–11.
Schwartz, R.E. et al. (2005) Defined conditions for development of functional hepatic cells from human embryonic stem cells. Stem Cells Dev. 14, 643–655.
Shirahashi, H. et al. (2004) Differentiation of human and mouse embryonic stem cells along a hepatocyte lineage. Cell Transplant. 13, 197–211.
Soderdahl, T. et al. (2007) Glutathione transferases in hepatocyte-like cells derived from human embryonic stem cells. Toxicol. In Vitro 21, 929–937.
Soto-Gutierrez, A. et al. (2006) Differentiation of human embryonic stem cells to hepatocytes using deleted variant of HGF and poly-amino-urethane-coated nonwoven polytetrafluoroethylene fabric. Cell Transplant. 15, 335–341.
D’Amour, K.A. et al. (2005) Efficient differentiation of human embryonic stem cells to definitive endoderm. Nat. Biotechnol. 23, 1534–1541.
Shiraki, N. et al. (2008) Differentiation of mouse and human embryonic stem cells into hepatic lineages. Genes Cells 13, 731–746.
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Lavon, N. (2010). Generation of Hepatocytes from Human Embryonic Stem Cells. In: Maurel, P. (eds) Hepatocytes. Methods in Molecular Biology, vol 640. Humana Press. https://doi.org/10.1007/978-1-60761-688-7_11
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DOI: https://doi.org/10.1007/978-1-60761-688-7_11
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