Establishment of the model of oxygen-glucose deprivation in vitro rat cortex and hippocampal neurons

Min Zheng, Chang-han Ou-Yang, Cui-yu Bao, Ji Zhao
Xianning College, Xianning City, Hubei Province, China

Our objective was to establish a model of oxygen-glucose deprivation in vitro rat cortex and hippocampal neurons. Cortex and hippocampal neurons cultured for 10∼12 days were exposed to Earle's solution with sodium dithionite(Na2S2O4)1mmol/l and free glucose in stand of original culture medium containing blood serum for 2, 4,8,12,24 and 36h respectively. 3-(4,5 - dimethylthiazol-2yl)—2,5—diphenylterazolium bromide(MTT) assay was applied to evaluate the neuronal vigor. A biochemical method was used to determined lactate dehydrogenase (LDH) activity. The rat cortex and hippocampal neurons vigor significantly decreased in a time-dependent manner after the original culture medium was replaced by Earle's solution with sodium dithionite 1mmol/l and free glucose for 2∼36h,and the neuron survival rate decreased to 25.4% and 24.2% respectively after oxygen-glucose deprivation for 36h. However, LDH release gradually increased in a time-dependent manner. There showed a strongly negative correlation for cortex or hippocampal neurons between the changes of LDH activity and neuronal survival rate (r = - 0.983, - 0.992, P<0.01). The model of oxygen-glucose deprivation in vitro rat cortex and hippocampal neurons was established successfully by applying Earle's solution with sodium dithionite 1mmol/l and free glucose.