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Purification of Hepatoblasts from Fetal Mouse Livers by Using a Temperature-Reversible Gelation Polymer and Their Application in Regenerative Medicine

Purification of Hepatoblasts and Their Application in Regenerative Medicine
  • Koichiro Yori
  • Toru Koike
  • Nobuyoshi Shiojiri
Conference paper
Part of the Animal Cell Technology: Basic & Applied Aspects book series (ANICELLTECH, volume 16)

Abstract

A convenient and efficient purification protocol for fetal mouse hepatoblasts and hepatocytes, which used a thermo-reversible gelation polymer (Mebiol), was established. When dispersed fetal mouse liver cells at E12.5 were cultured within the Mebiol gel, hepatoblasts rapidly aggregated into spheroids, and gave rise to hepatocytes expressing urea cycle enzymes on day 5. By contrast, nonparenchymal cells such as hepatic stellate cells and endothelial cells were dead within the gel. After culture for 3 or 5 days, fetal hepatoblast/hepatocyte aggregates could be easily separated from dead, single nonparenchymal cells by filtration. When E12.5 liver cells were cultured in a Mebiol gel containing basal laminar components, hepatoblasts gave rise to biliary epithelial cells forming cystic structures and expressing no hepatocyte marker. The culture and purification protocol for fetal liver progenitor cells may greatly contribute to regenerative medicine that aims at elucidating their growth and maturation mechanisms, and at developing novel hybrid-type artificial liver models and cell transplantation therapy.

Keywords

Hepatic Stellate Cell Lower Critical Solution Temperature Gelation Polymer Nonparenchymal Cell Biliary Epithelial Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Ikeda, T., Sawada, N., Satoh, M., and Mori, M. (1998) Induction of tyrosine aminotransferase of primary cultured rat hepatocytes depends on the organization of microtubules. J. Cell Physiol. 175: 41–49.CrossRefPubMedGoogle Scholar
  2. 2.
    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.CrossRefPubMedGoogle Scholar
  3. 3.
    Oertel, M., Menthena, A., Chen, Y., Teisner, B., Jensen, C., and Shafritz, D. (2008) Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver. Gastroenterology 134: 823–832.PubMedGoogle Scholar
  4. 4.
    Shiojiri, N. (1997) Development and differentiation of bile ducts in the mammalian liver. Microsc. Res. Tech. 39: 328–335.CrossRefPubMedGoogle Scholar
  5. 5.
    Suzuki, A., Sekiya, S., Büscher, D., Izpisúa Belmonte, J.C., and Taniguchi, H. (2008) Tbx3 controls the fate of hepatic progenitor cells in liver development by suppressing p19ARF expression. Development 135: 1589–1595.CrossRefPubMedGoogle Scholar
  6. 6.
    Tanimizu, N., Miyajima, A., and Mostov, K.E. (2007) Liver progenitor cells develop cholangiocyte-type epithelial polarity in three-dimensional culture. Mol. Cell Biol. 18: 1472–1479.CrossRefGoogle Scholar
  7. 7.
    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.CrossRefPubMedGoogle Scholar
  8. 8.
    Yoshioka, H., Mikami, M., Mori, Y., and Tsuchida, E. (1994) A synthetic hydrogel with thermoreversible gelation. II. Effect of added salts. J. Macromol. Sci. A31: 121–125.Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Department of BiologyShizuoka UniversitySuruga-ku, Shizuoka-CityJapan

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