Cryopreservation of monolayers of hepatocytes in a freezing medium containing 10% (v/v) dimethylsulfoxide, 90% (v/v) foetal calf serum retains cell morphology and viability, but cells lose up to 50% of their intracellular reduced glutathione. This is accompanied by a small increase in glutamate cysteine ligase expression in cryopreserved cultures, but glutathione synthetase expression is undetectable post-cryopreservation. Inclusion of ascorbic acid and alpha-tocopherol in the freezing medium improves maintenance of reduced glutathione content post-cryopreservation at 84% of the levels in non-cryopreserved monolayer cultures, but does not restore glutathione synthetase expression. The inability to synthesise reduced glutathione will mean that cryopreserved hepatocyte monolayers are more susceptible to toxic insults.
|Number of pages||5|
|Journal||Toxicology in Vitro|
|Publication status||Published - 2007|
- cryopreserved hepatocyte monolayers
- Reduced glutathione
- glutathione synthesis
- expression of glutathione synthetase
Stevenson, D. J., Morgan, C., McLellan, L. I., & Grant, M. H. (2007). Reduced glutathione levels and expression of the enzymes of glutathione synthesis in cryopreserved hepatocyte monolayer cultures. Toxicology in Vitro, 21(3), 527-532.