Use of a low-speed, iso-density percoll centrifugation method to increase the viability of isolated rat hepatocyte preparations

In Vitro Cell Dev Biol. 1986 Apr;22(4):201-11. doi: 10.1007/BF02623304.


A simple yet effective method (iso-density percoll centrifugation) has been developed for consistently preparing isolated rat liver parenchymal cells with over 98% initial viability. The method has been applied to cells isolated by a variety of collagenase digestion techniques. This procedure involves the low-speed centrifugation (50 X g) of the initial cell suspension through a percoll medium having a density of 1.06 g/ml and results in the separation of single and viable parenchymal cells from cell aggregates, debris, and nonparenchymal cells. The enriched parenchymal cells have been shown to be superior to untreated cells by a number of criteria including: preparation homogeneity, cell morphology, maintenance of cytochrome P-450, hormonal responsiveness (measured by the induction of tyrosine aminotransferase after treatment with glucagon or dexamethasone, or both), plasma membrane integrity (determined by both trypan blue exclusion and leakage of glutamic-oxaloacetic transaminase), and the DNA repair capability after treatment with benzo[a]pyrene or 2-acetylaminofluorene.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 2-Acetylaminofluorene / pharmacology
  • Animals
  • Benzo(a)pyrene / pharmacology
  • Cell Membrane / physiology
  • Cell Separation / methods*
  • Cell Survival
  • Centrifugation, Density Gradient
  • Cytochrome P-450 Enzyme System / metabolism
  • DNA / biosynthesis
  • DNA Repair
  • Dexamethasone / pharmacology
  • Enzyme Induction / drug effects
  • Glucagon / pharmacology
  • Liver / cytology*
  • Liver / metabolism
  • Male
  • Rats
  • Rats, Inbred Strains
  • Tyrosine Transaminase / biosynthesis


  • Benzo(a)pyrene
  • Dexamethasone
  • DNA
  • Glucagon
  • Cytochrome P-450 Enzyme System
  • 2-Acetylaminofluorene
  • Tyrosine Transaminase