Chapter 11. Isolation and Culture of Animal and Human Hepatocytes

  1. R. Ian Freshney and
  2. Mary G. Freshney
  1. Christiane Guguen-Guillouzo

Published Online: 28 APR 2002

DOI: 10.1002/0471221201.ch11

Culture of Epithelial Cells, Second Edition

Culture of Epithelial Cells, Second Edition

How to Cite

Guguen-Guillouzo, C. (2002) Isolation and Culture of Animal and Human Hepatocytes, in Culture of Epithelial Cells, Second Edition (eds R. I. Freshney and M. G. Freshney), John Wiley & Sons, Inc., New York, USA. doi: 10.1002/0471221201.ch11

Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

Author Information

  1. INSERM U522, Régulations des Equilibres Fonctionnels du Foie Normal et Pathologique, Hôpital Pontchaillou, av. de la Bataille, F-35033 Rennes, France

Publication History

  1. Published Online: 28 APR 2002
  2. Published Print: 12 APR 2002

Book Series:

  1. Culture of Specialized Cells

Book Series Editors:

  1. R. Ian Freshney

Series Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

ISBN Information

Print ISBN: 9780471401216

Online ISBN: 9780471221203

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Keywords:

  • liver epithelium;
  • hepatocytes;
  • epithelial cell culture;
  • collagenase perfusion;
  • cell cycle;
  • apoptosis;
  • rat liver epithelial cells;
  • RLEC;
  • myc;
  • jun;
  • EGF;
  • HGF;
  • matrix coating;
  • Matrigel;
  • EHS sarcoma;
  • differentiation;
  • immortalization

Summary

The chapter opens with a discussion of cell renewal and differentiation in the liver. A two-step collagenase perfusion technique is then described for the isolation of hepatocytes from a number of different species, with a separate protocol for obtaining hepatocytes from human liver. Purification of different subpopulations is also discussed. The control of entry into G1 of the cell cycle or the alternative entry into apoptosis has been examined in isolated hepatocytes involving alterations in the expression of c-myc and the jun family of genes. This is compared with equivalent steps in liver regeneration. Preservation of liver and isolated hepatocytes is described. Media and techniques used for extended culture are reviewed. The factors necessary for modulating cell proliferation, differentiation, and apoptosis are discussed. Matrix coating and communication with other rat liver epithelial cells (RLEC) are shown to be important in preserving the functional integrity of hepatocytes and protocols are described for establishment of RLEC and coculture. The chapter concludes with a discussion of differentiated cell lines and strategies for immortalization.