Advertisement

Isolation of Hepatic Progenitor Cells from the Galactosamine-Treated Rat Liver

  • Norihisa IchinoheEmail author
  • Junko Kon
  • Toshihiro MitakaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 826)

Abstract

Oval cells and small hepatocytes (SHs) are well known as hepatic stem/progenitor cells. However, the relationship between the oval cells and SHs in liver regeneration is not well understood. To resolve this issue, we established a technique to selectively separate oval cells and SHs. In the injured rat liver, oval cells and SHs transiently appear in the initial period of liver regeneration. Thy1+ and CD44+ cells are candidates for markers of oval cells and SHs, respectively. In this chapter, the methods for sorting and culture of the cells are described in detail.

Key words

Oval cells Small hepatocytes Thy1 CD44 Liver Stem/progenitor cells Galactosamine 

Notes

Acknowledgments

We thank Ms. Minako Kuwano and Ms. Yumiko Tsukamoto for technical assistance. We also thank Mr. Kim Barrymore for help with the manuscript. This work was supported by Grants-in-Aid for Scientific Research (for N. Ichinohe, J. Kon, and for T. Mitaka) and the program for developing the supporting system for upgrading education and research (for T. Mitaka) from the Ministry of Education, Culture, Sports, Science, and Technology, the Science and Technology Incubation Program in Advanced Regions from the Japan Science and Technology Agency (for T. Mitaka), Research on Advanced Medical Technology from the Ministry of Health, Labor, and Welfare, and Labor Sciences Research Grants (for T. Mitaka), and the Suhara Memorial Foundation (for T. Mitaka).

References

  1. 1.
    Sell S: The role of progenitor cells in repair of liver injury and in liver transplantation. Wound Rep Reg 2001, 9: 467–482CrossRefGoogle Scholar
  2. 2.
    Forbes S, Vig P, Poulsom R, Thomas H, Alison M. Hepatic stem cells: J Pathol 2002, 197:510–518PubMedGoogle Scholar
  3. 3.
    Fausto N, Campbell JS: The role of hepatocytes and oval cells in liver regeneration and repopulation. Mech Dev 2003, 120:117–130PubMedCrossRefGoogle Scholar
  4. 4.
    Roskams TA, Libbrecht L, Desmet VJ: Progenitor cells in diseased human liver. Semin Liver Dis 2003, 23:385–396PubMedCrossRefGoogle Scholar
  5. 5.
    Knight B, Matthews VB, Olynyk JK, Yeoh GC: Jekyll and Hyde: evolving perspectives on the function and potential of the adult liver progenitor (oval) cell. BioEssays 2005, 27:1192–1202PubMedCrossRefGoogle Scholar
  6. 6.
    Walkup MH, Gerber DA: Hepatic stem cells: in search of. Stem Cells 2006, 24:1833–1840PubMedCrossRefGoogle Scholar
  7. 7.
    Farber E: Similarities in the sequence of early histological changes induced in the liver of the rat by ethionine, 2-acetylaminofluorene, and 3’-methy-4-dimethylaminoazobenzene. Cancer Res 1956, 16:142–149PubMedGoogle Scholar
  8. 8.
    Petersen BE, Goff JP, Greenberger JS, Michalopoulos GK: Hepatic oval cells express the hematopoietic stem cell marker Thy-1 in the rat. Hepatology 1998, 27:433–45PubMedCrossRefGoogle Scholar
  9. 9.
    Mitaka T, Kojima T, Mizuguchi T, Mochizuki Y: Growth and maturation of small hepatocytes isolated from adult liver. Biochem Biophys Res Commun 1995, 214:310–317PubMedCrossRefGoogle Scholar
  10. 10.
    Mitaka T, Sato F, Mizuguchi T, Yokono T, Mochizuki Y: Reconstruction of hepatic organoid by rat small hepatocytes and hepatic nonparenchymal cells. Hepatology 1999, 29:111–125PubMedCrossRefGoogle Scholar
  11. 11.
    Sugimoto S, Mitaka T, Ikeda S, Harada K, Ikai I, Yamaoka Y, Mochizuki Y: Morphological changes induced by extracellular matrix are correlated with maturation of rat small hepatocytes. J Cell Biochem 2002, 1:16–28CrossRefGoogle Scholar
  12. 12.
    Mitaka T, Ooe H. Drug Metabolism Reviews focusing on drug transporter interactions in the liver: characterization of hepatic-organoid cultures. Drug Metab Rev 2010, 42:472–481Google Scholar
  13. 13.
    Kon J, Ooe H, Oshima H, Kikkawa Y, Mitaka T: Expression of CD44 in rat hepatic progenitor cells. J Hepatol 2006, 45:90–98PubMedCrossRefGoogle Scholar
  14. 14.
    Kon J, Ichinohe N, Ooe H, Chen Q, Sasaki K, Mitaka T. Thy1-positive cells have bipotential ability to differentiate into hepatocytes and biliary epithelial cells in galactosamine-induced rat liver regeneration. Am J Pathol 2009, 175:2362–2371PubMedCrossRefGoogle Scholar
  15. 15.
    Chen Q, Kon J, Ooe H, Sasaki K, Mitaka T: Selective proliferation of rat hepatocyte progenitor cells in serum-free culture. Nat Protoc 2007, 2:1197–1205PubMedCrossRefGoogle Scholar
  16. 16.
    Sasaki K, Kon J, Mizuguchu T, Chen Q, Ooe H, Oshima H, Hirata K, Mitaka T: Proliferation of hepatocyte progenitor cells isolated from adult human livers in serum-free medium. Cell Transplant. 2008, 17:1221–1230PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Tissue Development and Regeneration, Research Institute for Frontier MedicineSapporo Medical University School of MedicineSapporoJapan

Personalised recommendations