A Modified Protocol for the Isolation of Primary Human Hepatocytes with Improved Viability and Function from Normal and Diseased Human Liver

Methods Mol Biol. 2017;1506:61-73. doi: 10.1007/978-1-4939-6506-9_4.


Successful hepatocyte isolation is critical for continued development of cellular transplantation. However, most tissue available for research is from diseased liver and the results of hepatocyte isolation from such tissue are inferior compared to normal tissue. Here we describe a modified method, combining the use of Liberase and N-acetylcysteine (NAC), for the isolation of primary human hepatocytes with high viability from normal and diseased liver.

Keywords: Hepatocyte isolation; Hepatocyte viability; Human hepatocyte; Liberase; N-Acetylcysteine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / pharmacology
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Survival / drug effects
  • Cell Transplantation / methods*
  • Cells, Cultured
  • Collagenases / pharmacology
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism
  • Hepatocytes / transplantation*
  • Humans
  • Liver / cytology*
  • Liver Diseases / surgery*
  • Primary Cell Culture
  • Thermolysin / pharmacology
  • Tissue Donors


  • Collagenases
  • Liberase
  • Thermolysin
  • Acetylcysteine