Abstract
To investigate cell–cell interactions during mammalian liver development, it is essential to separate hepatoblasts (fetal liver progenitor cells) from nonparenchymal cells, including stellate cells, endothelial cells, and hemopoietic cells. Various factors, which may be produced by nonparenchymal cells, could be assayed for their effects on the growth and maturation of separated hepatoblasts. The protocol using immunomagnetic beads coated with anti-mouse E-cadherin antibody is described for efficient isolation of hepatoblasts from cell suspensions of fetal mouse livers. The purity and recovery rate are larger than 95% and approximately 30%, respectively. The protocol may be useful for various studies focusing on the fetal liver progenitor cells.
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
Shiojiri, N. (1997) Development and differentiation of bile ducts in the mammalian liver. Microsc Res Tech 39, 328–335.
Lemaigre, F. P. (2009) Mechanisms of liver development: concepts for understanding liver disorders and design of novel therapies. Gastroenterology 137, 62–79.
Nitou, M., Sugiyama, Y., Ishikawa, K. and Shiojiri, N. (2002) Purification of fetal mouse hepatoblasts by magnetic beads coated with monoclonal anti-E-cadherin antibodies and their in vitro culture. Exp Cell Res 279, 330–343.
Yasuchika, K., Hirose, T., Fujii, H., Oe, S., Hasegawa, K., Fujikawa, T., Azuma, H. and Yamaoka, Y. (2002) Establishment of a highly efficient gene transfer system for mouse fetal hepatic progenitor cells. Hepatology 36, 1488–1497.
Tanimizu, N., Nishikawa, M., Saito, H., Tsujimura, T. and Miyajima, A. (2003) Isolation of hepatoblasts based on the expression of Dlk/Pref-1. J Cell Sci 116, 1775–1786.
Hirose, Y., Itoh, T. and Miyajima, A. (2009) Hedgehog signal activation coordinates proliferation and differentiation of fetal liver progenitor cells. Exp Cell Res 315, 2648–2657.
Murphy, S. J., Watt, D. J. and Jones, G. E. (1992) An evaluation of cell separation techniques in a model mixed cell population. J Cell Sci 102, 789–798.
Acknowledgments
This work was supported in part by Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science, and Technology, the Japanese Government.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Shiojiri, N., Nitou, M. (2012). Purification and Culture of Fetal Mouse Hepatoblasts that Are Precursors of Mature Hepatocytes and Biliary Epithelial Cells. In: Ochiya, T. (eds) Liver Stem Cells. Methods in Molecular Biology, vol 826. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-468-1_1
Download citation
DOI: https://doi.org/10.1007/978-1-61779-468-1_1
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-61779-467-4
Online ISBN: 978-1-61779-468-1
eBook Packages: Springer Protocols