Stevenson David J, Morgan Caroline, McLellan Lesley I, Helen Grant M
Bioengineering Unit, University of Strathclyde, Wolfson Centre, 106 Rottenrow, Glasgow G4 0NW, UK.
Toxicol In Vitro. 2007 Apr;21(3):527-32. doi: 10.1016/j.tiv.2006.11.005. Epub 2006 Nov 21.
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.
在含有10%（体积/体积）二甲基亚砜和90%（体积/体积）胎牛血清的冷冻培养基中对单层肝细胞进行冷冻保存，可保持细胞形态和活力，但细胞会损失高达50%的细胞内还原型谷胱甘肽。这伴随着冷冻保存培养物中谷氨酸半胱氨酸连接酶表达的小幅增加，但冷冻保存后未检测到谷胱甘肽合成酶的表达。在冷冻培养基中加入抗坏血酸和α-生育酚可将冷冻保存后还原型谷胱甘肽含量维持在非冷冻保存单层培养物水平的84%，但无法恢复谷胱甘肽合成酶的表达。无法合成还原型谷胱甘肽意味着冷冻保存的单层肝细胞更容易受到毒性损伤。
