DARU Journal of Pharmaceutical Sciences 2009. 17(3):187-191.

Effect of dithiotheritol on viability of cryopreserved rat hepatocytes
A Jamshidzadeh, M Hosseinaghdaie, H Niknahad, M Nematizadeh


Background: Prolonged storage of cryopreserved isolated hepatocytes is now possible and such cells have been used with success for drug metabolism and toxicity studies. Quality of the cells before cryopreservation is important for viability and function after freezing.

Methods: In this study, fresh rat hepatocytes were incubated with the thiol-containing compounds N-acetylcysteine (NAC), dithiotheritol (DTT) and fructose as ATP supplier, at incubation times 1 and 3 hrs. The preincubated hepatocytes cryopreserved for 24 hour, and 1 and 3 months. Hepatocytes, viability were determined immediately postthaw by Trypan Blue exclusion.

Results: Fructose preincubation improved the viability of hepatocytes at a concentration of 300 mM. Preincubation with DTT (50, 100 and 200 μM) prior to cryopreservation had beneficial effects on viability of hepatocytes. The postthaw viability of hepatocytes preincubated with NAC was uniformly poor.

Conclusion: Preincubation with DTT, improved GSH levels before freezing which could be responsible for the reduction in membrane damage during cryopreservation. It may be concluded that the intracellular


Cryopreservation, Dithiotheritol, Hepatocytes,

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