Abstract
Investigations on the biology of hepatitis C virus (HCV) have been hampered by the lack of small animal models. Efforts have therefore been directed to designing practical and robust cellular models of human origin able to support HCV replication and production in a reproducible and physiologically pertinent manner. Different systems have been constructed based on hepatoma or other cell lines, sub-genomic and genomic replicons, productive replicons, and immortalized hepatocytes. Although these models are practical for high-throughput screenings, they present several drawbacks related to the nature of the virions and the fact that the cells are not differentiated. Adult primary human hepatocytes infected with natural serum-derived HCV virions represent the model that most closely mimics the physiological situation. This chapter describes our experience with this culture model.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chevaliez, S. and Pawlotsky, J.M. (2007) Hepatitis C virus: virology, diagnosis and management of antiviral therapy. World J. Gastroenterol. 13, 2461–2466.
Lavanchy, D. and Gavinio, P. (2000) Hepatitis C. Can. J. Gastroenterol. 14 Suppl B, 67B–76B.
Chevaliez, S. and Pawlotsky, J.M. (2007) Interferon-based therapy of hepatitis C. Adv. Drug Deliv. Rev. 59(12), 1222–1241.
Bartenschlager, R. (2005) The hepatitis C virus replicon system: from basic research to clinical application. J. Hepatol. 43, 210–216.
Lazaro, C.A., Chang, M., Tang, W. et al. (2007) Hepatitis C virus replication in transfected and serum-infected cultured human fetal hepatocytes. Am. J. Pathol. 170, 478–489.
Kato, T., Date, T., Murayam, A. et al. (2006) Cell culture and infection system for hepatitis C virus. Nat. Protoc. 1, 2334–2339.
Lindenbach, B.D., Evans, M.J., Syder, A.J. et al. (2005) Complete replication of hepatitis C virus in cell culture. Science 309, 623–626.
Zhong, J., Gastaminza, P., Cheng, G. et al. (2005) Robust hepatitis C virus infection in vitro. Proc. Natl. Acad. Sci. USA 102, 9294–9299.
Fournier, C., Sureau, C., Coste, J. et al. (1998) In vitro infection of adult normal human hepatocytes in primary culture by hepatitis C virus. J. Gen. Virol. 79, 2367–2374.
Gondeau, C., Pichard-Garcia, L., and Maurel, P. (2009) Cellular models for the screening and development of anti-hepatitis C virus agents. Pharmacol. Ther. 124, 1–22.
Buck, M. (2008) Direct infection and replication of naturally occurring hepatitis C virus genotypes 1, 2, 3 and 4 in normal human hepatocyte cultures. PLoS ONE 3, e2660.
Rumin, S., Berthillon, P., Tanaka, E. et al. (1999) Dynamic analysis of hepatitis C virus replication and quasispecies selection in long-term cultures of adult human hepatocytes infected in vitro. J. Gen. Virol. 80, 3007–3018.
Carriere, M., Pene, V., Breiman, A. et al. (2007) A novel, sensitive, and specific RT-PCR technique for quantitation of hepatitis C virus replication. J. Med. Virol. 79, 155–160.
Chong, T.W., Smith, R.L., Hughes, M.G. et al. (2006) Primary human hepatocytes in spheroid formation to study hepatitis C infection. J. Surg. Res. 130, 52–57.
Castet, V., Fournier, C., Soulier, A. et al. (2002) Alpha interferon inhibits hepatitis C virus replication in primary human hepatocytes infected in vitro. J. Virol. 76, 8189–8199.
Molina, S., Castet, V., Pichard-Garcia, L. et al. (2008) Serum-derived hepatitis C virus infection of primary human hepatocytes is tetraspanin CD81 dependent. J. Virol. 82, 569–574.
Molina, S., Castet, V., Fournier-Wirth, C. et al. (2007) The low-density lipoprotein receptor plays a role in the infection of primary human hepatocytes by hepatitis C virus. J. Hepatol. 46, 411–419.
Codran, A., Royer, C., Jaeck, D. et al. (2006) Entry of hepatitis C virus pseudotypes into primary human hepatocytes by clathrin-dependent endocytosis. J. Gen. Virol. 87, 2583–2593.
Kato, T., Date, T., Miyamoto, M. et al. (2003) Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. Gastroenterology 125, 1808–1817.
Rouille, Y., Helle, F., Delgrange, D. et al. (2006) Subcellular localization of hepatitis C virus structural proteins in a cell culture system that efficiently replicates the virus. J. Virol. 80, 2832–2841.
Abe, K., Nozaki, A., Tamura, K. et al. (2007) Tandem repeats of lactoferrin-derived anti-hepatitis C virus peptide enhance antiviral activity in cultured human hepatocytes. Microbiol. Immunol. 51, 117–125.
Ikeda, M., Sugiyama, K., Tanaka, T. et al. (1998) Lactoferrin markedly inhibits hepatitis C virus infection in cultured human hepatocytes. Biochem. Biophys. Res. Commun. 245, 549–553.
Acknowledgments
Part of the work described has been supported by Roche Ltd., Neilly France.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Pichard-Garcia, L. et al. (2010). Use of Human Hepatocytes to Investigate HCV Infection. In: Maurel, P. (eds) Hepatocytes. Methods in Molecular Biology, vol 640. Humana Press. https://doi.org/10.1007/978-1-60761-688-7_24
Download citation
DOI: https://doi.org/10.1007/978-1-60761-688-7_24
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60761-687-0
Online ISBN: 978-1-60761-688-7
eBook Packages: Springer Protocols