An improved method of making oxygen-glucose deprivation model of hippocampal slice in vitro in neonatal rats

Objective:To introduce an economical and pragmatic method of establishing oxygen-glucose deprivation model of hippocampal slice in neonatal rats in vitro,Methods:Hippocampal slices(400μm)from 8~10-day-old rats are transferred into an incubator with insert of Millicell membrance and incubated for 7 d,14 d,20 d and 30 d,respectively.The growing state of the hippocampal slices were observed under the convened microscope,and its ability to absorb propidiumiodide (PI) was detected under fluorescent microscope after the hippocampal slice was stained by PI,and the state of the hippocampal slices at different time after aerating Nitrogen gas was observed. Results: ① No PI labelled cells were found after 7 d and 14 d. However,15% PI positive cells were found after 30 d. ②The longer in Nitrogen gas,the more apoptosis cells. ③Intensive fluorescence was observed in the CA1,CA3 and DG areas of the hippocampal slice after aerating Nitrogen gas for 1.5h. Conclusion:The hippocampal slice can be alive for about 30 days.Stable oxygen-glucose deprivation model of the hippocampl slice in vitro was successfully established after aerating Nitrogen gas for 1.5 h.