Metabolic activity of fresh and cryopreserved cynomolgus monkey (Macaca fascicularis) hepatocytes

Xenobiotica. 2000 Jul;30(7):665-81. doi: 10.1080/00498250050077986.


1. The effect of cryopreservation on the metabolic capacity of monkey hepatocytes over 4 h in suspension and 24 h in culture was determined. Hepatocytes were diluted in a buffer containing 10% DMSO and frozen in a computer-controlled chamber. 2. Initial ethoxyresorufin and ethoxycoumarin O-deethylase (ECOD) activities were the same in fresh and cryopreserved (CP) hepatocytes. ECOD activity in suspensions declined over 4 h but was the same in fresh and CP hepatocytes. 3. The formation of testosterone hydroxy (OHT) metabolites (namely 6beta-OHT, 2beta-OHT, 16beta-OHT, 16alpha-OHT, 15beta-OHT, 2alpha-OHT and 6beta-OHT) was unaffected by cryopreservation. The loss of OHT activities over 4 h in CP and fresh whole cell suspensions was attributed to a loss of cofactor. CP hepatocyte cultures had equivalent OHT activities to freshly isolated hepatocytes. 4. Initial UDP-glucuronyltransferase (UGT) activities, using the substrates 4-methylumbelliferone, ethoxycoumarin and hydroxycoumarin, were equivalent in fresh and CP whole hepatocytes. At later times, UGT activity was lower in CP than fresh hepatocytes but this was due to a loss of UDPGA. Initial sulphotransferase (SULT) activities, using the substrates 2-naphthol, ethoxycoumarin and hydroxycoumarin, were equivalent in fresh and CP hepatocytes. SULT activities were less stable than UGT activities but were the same in fresh and CP hepatocytes throughout the 4-h incubation. 5. Initial glutathione S-transferase activities (using 1-chloro-2,4-dinitrobenzene) were the same in fresh and CP hepatocytes and both did not decrease over 4 h. 6. CP monkey hepatocytes are a useful model for metabolic and cytotoxicity studies. These cells can be can be used either in suspension or in culture.

Publication types

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

MeSH terms

  • Animals
  • Aryl Hydrocarbon Hydroxylases*
  • Cell Adhesion
  • Cell Separation
  • Cell Survival
  • Cells, Cultured
  • Cryopreservation*
  • Cytochrome P-450 Enzyme System / metabolism
  • Glutathione / metabolism
  • Hepatocytes / enzymology
  • Hepatocytes / metabolism*
  • In Vitro Techniques
  • L-Lactate Dehydrogenase / metabolism
  • Macaca fascicularis
  • Male
  • Steroid Hydroxylases / metabolism


  • Cytochrome P-450 Enzyme System
  • L-Lactate Dehydrogenase
  • Steroid Hydroxylases
  • Aryl Hydrocarbon Hydroxylases
  • testosterone 7-alpha-hydroxylase, hamster
  • Glutathione