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

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